Stem Cell Clinic

CardiAMP Cell Therapy System – biocardia.com

By Uncategorized

CardiAMP Cell Therapy System The investigational CardiAMP Therapy is designed to be a comprehensive biotherapeutic heart failure solution, incorporating:

Day 1: Mini bone marrow aspirate (~ 1 tablespoon) of patients bone marrow cells from their hip bone. Aspirate sent to BioCardia CLIA laboratory partner. BioCardias proprietary molecular diagnostic assay identifies patients with potent cell characteristics using an In Vitro Diagnostic Multi Indexed Assay.

Day 3 or after: If assay positive, patient presents for 1-hour treatment in cardiac catheterization lab:

Small bone marrow aspirate ( ~4 tablespoons) ~15 minutes Cells minimally processed and concentrated into proprietary dosage Cells delivered using proprietary Helix Transendocardial Delivery System Patient discharged same day or after overnight stay

In both thePhase I clinical trialof 20 patients (the TABMMI trial) and Phase II randomized placebo-controlled trial of 30 patients (the TACHFT-BMC Trial) CardiAMP cells demonstrated an excellent safety profile in heart failure patients treated at two dosages. There have been no incidences of treatment-emergent major adverse cardiac events. The Phase II results from TACHFT-BMC support efficacy relative to patient quality of life and functional capacity, which was shown to be both statistically and clinically significant.The FDA has approved us to begin enrollment of patients in our CardiAMP Phase 3 pivotal, pre-commercial trial using a patient's owncells, administered in the cardiac catheterization lab directly into the heart muscle in cases of seriousheart failure following a priorheart attack.The trial is expected to beginin approved centers in late 2016 or early 2017.

Raval AN, Cook TD, Duckers HJ, Johnston PV, Traverse JH, Abraham WT, Altman PA, Pepine CJ. The CardiAMP Heart Failure trial: A randomized controlled pivotal trial of high-dose autologous bone marrow mononuclear cells using the CardiAMP cell therapy system in patients with postmyocardial infarction heart failure: Trial rationale and study design,American Heart Journal, April 2018.

Mitsutake Y, Pyum WB, Rouy D, et al. Improvement of local cell delivery using Helix Transendocardial Delivery Catheter in a porcine heart.Int Heart J. 2017.

Heldman AW, Difede DL, Fishman JE, Zambrano JP, Trachtenberg BH, Karantalis V, Mushtaq M, Williams AR, Suncion VY, McNiece IK, Ghersin E, Soto V, Lopera G, Miki R, Willens H, Hendel R, Mitrani R, Pattany P, Feigenbaum G, Oskouei B, Byrnes J, Lowery MH, Sierra J, Pujol MV, Delgado C, Gonzalez PJ, Rodriguez JE, Bagno LL, Rouy D, Altman P, Foo CW, da Silva J, Anderson E, Schwarz R, Mendizabal A, Hare JM. Transendocardial mesenchymal stem cells and mononuclear bone marrow cells for ischemic cardiomyopathy: The TAC-HFT randomized trial.JAMA. 2013

Wong Po Foo C, Ikeno F, Altman P, Rouy D. Quantifying therapeutic cell retention in the heart to compare three routes of local delivery: Transendocardial intramyocardial injection, transepicardial intramyocardial injection, and intracoronary artery infusion.8th International Conference on Cell Therapy for Cardiovascular Disease. 2013

de la Fuente LM, Stertzer SH, Argentieri J, Penaloza E, Koziner B, Rouy D, Altman PA. Transendocardial autologous bone marrow in myocardial infarction induced heart failure, two-year follow-up in an open-label phase i safety study (the tabmmi study).EuroIntervention : journal of EuroPCR in collaboration with theWorking Group on Interventional Cardiology of the European Society of Cardiology. 2011;7:805-812.

de la Fuente LM, Stertzer SH, Argentieri J, Penaloza E, Miano J, Koziner B, Bilos C, Altman PA. Transendocardial autologous bone marrow in chronic myocardial infarction using a helical needle catheter: 1-year follow-up in an open-label, nonrandomized, single-center pilot study (the tabmmi study).Am Heart J. 2007;154:79 e71-77

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CardiAMP Cell Therapy System - biocardia.com

Mosaic (genetics) – Wikipedia

By Somatic Stem Cells

In genetics, a mosaic, or mosaicism, involves the presence of two or more populations of cells with different genotypes in one individual who has developed from a single fertilized egg.[1][2] Mosaicism has been reported to be present in as high as 70% of cleavage stage embryos and 90% of blastocyst-stage embryos derived from in vitro fertilization.[3]

Genetic mosaicism can result from many different mechanisms including chromosome non-disjunction, anaphase lag, and endoreplication.[3] Anaphase lagging is the most common way by which mosaicism arises in the preimplantation embryo.[3] Mosaicism can also result from a mutation in one cell during development in which the mutation is passed on to only its daughter cells. Therefore, the mutation is only going to be present in a fraction of the adult cells.[2]

Genetic mosaics may often be confused with chimerism, in which two or more genotypes arise in one individual similarly to mosaicism. However, in chimerism the two genotypes arise from the fusion of more than one fertilized zygote in the early stages of embryonic development, rather than from a mutation or chromosome loss.

Different types of mosaicism exist, such as gonadal mosaicism (restricted to the gametes) or somatic mosaicism.

Somatic mosaicism occurs when the somatic cells of the body are of more than one genotype. In the more common mosaics, different genotypes arise from a single fertilized egg cell, due to mitotic errors at first or later cleavages.

In rare cases, intersex conditions can be caused by mosaicism where some cells in the body have XX and others XY chromosomes (46, XX/XY).[4][5] In the fruit fly Drosophila melanogaster, where a fly possessing two X chromosomes is a female and a fly possessing a single X chromosome is a sterile male, a loss of an X chromosome early in embryonic development can result in sexual mosaics, or gynandropmorphs.[6][7] Likewise, a loss of the Y chromosome can result in XY/X mosaic males.[8]

The most common form of mosaicism found through prenatal diagnosis involves trisomies. Although most forms of trisomy are due to problems in meiosis and affect all cells of the organism, there are cases where the trisomy occurs in only a selection of the cells. This may be caused by a nondisjunction event in an early mitosis, resulting in a loss of a chromosome from some trisomic cells.[9] Generally this leads to a milder phenotype than in non-mosaic patients with the same disorder.

An example of this is one of the milder forms of Klinefelter syndrome, called 46/47 XY/XXY mosaic wherein some of the patient's cells contain XY chromosomes, and some contain XXY chromosomes. The 46/47 annotation indicates that the XY cells have the normal number of 46 total chromosomes, and the XXY cells have a total of 47 chromosomes.

Around 30% of Turner's syndrome cases demonstrate mosaicism, while complete monosomy (45, X) occurs in about 5060% of cases.

But mosaicism need not necessarily be deleterious. Revertant somatic mosaicism is a rare recombination event in which there is a spontaneous correction of a mutant, pathogenic allele.[10] In revertant mosaicism, the healthy tissue formed by mitotic recombination can outcompete the original, surrounding mutant cells in tissues like blood and epithelia that regenerate often.[10] In the skin disorder ichthyosis with confetti, normal skin spots appear early in life and increase in number and size over time.[10]

Other endogenous factors can also lead to mosaicism including mobile elements, DNA polymerase slippage, and unbalanced chromosomal segregation.[11] Exogenous factors include nicotine and UV radiation.[11] Somatic mosaics have been created in Drosophila using Xray treatment and the use of irradiation to induce somatic mutation has been a useful technique in the study of genetics.[12]

True mosaicism should not be mistaken for the phenomenon of Xinactivation, where all cells in an organism have the same genotype, but a different copy of the X chromosome is expressed in different cells. The latter is the case in normal (XX) female mammals, although it is not always visible from the phenotype (like it is in calico cats). However, all multicellular organisms are likely to be somatic mosaics to some extent.[13]

Somatic mutation leading to mosaicism is prevalent in the beginning and end stages of human life.[11] Somatic mosaics are common in embryogenesis due to retrotransposition of L1 and Alu transposable elements.[11] In early development, DNA from undifferentiated cell types may be more susceptible to mobile element invasion due to long, un-methylated regions in the genome.[11] Further, the accumulation of DNA copy errors and damage over a lifetime lead to greater occurrences of mosaic tissues in aging humans. As our longevity has increased dramatically over the last century, our genome may not have had time to adapt to cumulative effects of mutagenesis.[11] Thus, cancer research has shown that somatic mutations are increasingly present throughout a lifetime and are responsible for most leukemia, lymphomas, and solid tumors.[14]

Genomic mosaiscism arises in developing and in adult brain cells leading to diverse, seemingly random, genomic changes.[15] A frequent type of neuronal genomic mosaicism is copy number variation. Possible sources of such variation were suggested to be incorrect repair of DNA damages and somatic recombination.[15][16]

One basic mechanism which can produce mosaic tissue is mitotic recombination or somatic crossover. It was first discovered by Curt Stern in Drosophila in 1936. The amount of tissue which is mosaic depends on where in the tree of cell division the exchange takes place. A phenotypic character called "Twin Spot" seen in Drosophila is a result of mitotic recombination. However, it also depends on the allelic status of the genes undergoing recombination. Twin spot occurs only if the heterozygous genes are linked in repulsion i.e. trans phase. The recombination needs to occur between the centromere the adjacent gene. This gives an appearance of yellow patches on the wild type background in Drosophila. another example of mitotic recombination is the Bloom's syndrome which happens due to the mutation in the blm gene. The resulting BLM protein is defective. the defect in RecQ an helicase facilitates the defective unwinding of DNA during replication and is thus associated with the occurrence of this disease.[17][18]

Germline or gonadal mosaicism is a special form of mosaicism, where some gametesi.e., sperm or oocytescarry a mutation, but the rest are normal.[19][20]

The cause is usually a mutation that occurred in an early stem cell that gave rise to all or part of the gametes.

This can cause only some offspring to be affected, even for a dominant disease.

Genetic mosaics can be extraordinarily useful in the study of biological systems, and can be created intentionally in many model organisms in a variety of ways. They often allow for the study of genes that are important for very early events in development, making it otherwise difficult to obtain adult organisms in which later effects would be apparent. Furthermore, they can be used to determine the tissue or cell type in which a given gene is required and to determine whether a gene is cell autonomous. That is, whether or not the gene acts solely within the cell of that genotype, or if it affects the entire organism of neighboring cells which do not themselves contain that genotype.

The earliest examples of this involved transplantation experiments (technically creating chimeras) where cells from a blastula stage embryo from one genetic background are aspirated out and injected into a blastula stage embryo of a different genetic background.

Genetic mosaics are a particularly powerful tool when used in the commonly studied fruit fly, where specially-selected strains frequently lose an X[7] or a Y[8] chromosome in one of the first embryonic cell divisions. These mosaics can then be used to analyze such things as courtship behavior,[7] female sexual attraction,[21] and the autonomy or non-autonomy of particular genes.

Genetic mosaics can also be created through mitotic recombination. Such mosaics were originally created by irradiating flies heterozygous for a particular allele with X-rays, inducing double-strand DNA breaks which, when repaired, could result in a cell homozygous for one of the two alleles. After further rounds of replication, this cell would result in a patch, or "clone" of cells mutant for the allele being studied.

More recently the use of a transgene incorporated into the Drosophila genome has made the system far more flexible. The flip recombinase (or FLP) is a gene from the commonly studied yeast Saccharomyces cerevisiae which recognizes "flip recombinase target" (FRT) sites, which are short sequences of DNA, and induces recombination between them. FRT sites have been inserted transgenically near the centromere of each chromosome arm of Drosophila melanogaster. The FLP gene can then be induced selectively, commonly using either the heat shock promoter or the GAL4/UAS system. The resulting clones can be identified either negatively or positively.

In negatively marked clones the fly is transheterozygous for a gene encoding a visible marker (commonly the green fluorescent protein or GFP) and an allele of a gene to be studied (both on chromosomes bearing FRT sites). After induction of FLP expression, cells that undergo recombination will have progeny that are homozygous for either the marker or the allele being studied. Therefore, the cells that do not carry the marker (which are dark) can be identified as carrying a mutation.

It is sometimes inconvenient to use negatively marked clones, especially when generating very small patches of cells, where it is more difficult to see a dark spot on a bright background than a bright spot on a dark background. It is possible to create positively marked clones using the so-called MARCM ("mosaic analysis with a repressible cell marker", pronounced [mark-em]) system, developed by Liqun Luo, a professor at Stanford University, and his post-doc Tzumin Lee who now leads a group at Janelia Farm Research Campus. This system builds on the GAL4/UAS system, which is used to express GFP in specific cells. However a globally expressed GAL80 gene is used to repress the action of GAL4, preventing the expression of GFP. Instead of using GFP to mark the wild-type chromosome as above, GAL80 serves this purpose, so that when it is removed by mitotic recombination, GAL4 is allowed to function, and GFP turns on. This results in the cells of interest being marked brightly in a dark background.[22]

In 1929, Alfred Sturtevant studied mosaicism in Drosophila.[6] A few years later, In the 1930s, Curt Stern demonstrated that genetic recombination, normal in meiosis, can also take place in mitosis.[23][24] When it does, it results in somatic (body) mosaics. These are organisms which contain two or more genetically distinct types of tissue.[25] The term "somatic mosaicism" was used by C.W. Cotterman in 1956 in his seminal paper on antigenic variation.[11]

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Mosaic (genetics) - Wikipedia

Platelet-Rich Plasma Injections for knee osteoarthritis …

By Platelet Rich Plasma Injections

Marc Darrow MD, JD. Thank you for reading my article. You can ask me your questions using the contact form below.

While we do offer stem cell therapy for knee osteoarthritis, the focus of this article will be onPlatelet-Rich Plasma Therapy. We receive many emails which ask which is the better treatment for me, stem cells or PRP? That answer comes best after a physical examination and when we have the opportunity to sit down together and discuss what are your goals of treatment. Goals of treatment would be different for a mountain climber than a stair climber at home.

In the same regard, we get many emails that ask us if PRP is better thanhyaluronic acid injections or cortisone injection. Many of these people already had cortisone andhyaluronic acid injections. They are somewhat skeptical because these treatments did not help them as much as they thought they would.

In our experience, we have found PRP injections to be the superior treatment. In fact we have submitted our findings for peer-review and upcoming medical journal publication. Supporting our view most recently is a2019 study in the medical journalOrthopade(1) which found thatIntra-articular PRP injections into the knee for symptomatic early stages of knee osteoarthritis are a valid treatment option. The clinical efficacy of Intra-articular PRP injections is comparable to that of the Intra-articular-hyaluronic acid injections and Intra-articular cortisone after 3 months, HOWEVER, the long-term effectiveness of PRP injections is superior to hyaluronic acid and cortisone.

In this article we will discuss research on grade 1 to 3 knee osteoarthritis and PRP treatments. When a new patient comes into our office for a consultation for their knee osteoarthritis, we do a careful assessment of the patient and then make recommendations. Sometimes, the lack of range of motion in this patients knee and other factors lead us to a recommendation of stem cell injections. This recommendation is based on a realistic expectation of what both treatments may offer. For some, having the PRP only may not offer the healing that they hope to achieve.

Doctors at the worlds leading medical universities and hospitals are showing that PRP can regenerate damaged knee cartilage and meniscus in patients suffering from knee osteoarthritis and PRP can also enhance healing after knee ligament reconstruction.

An October 2018 study in the journal Current reviews in musculoskeletal medicine(2) says this:

Recent research into the applications ofPRPfor knee osteoarthritis has further indicated both the efficacy and safety ofPRPtreatment. Although research has shown a tendency toward better efficacy at earlier stages ofosteoarthritis, evidence exists to indicate positive effects at all stages of osteoarthritis. In summary, since knee osteoarthritis is an extremely prevalent condition that can be a challenge to treat, it is imperative that safe and effective nonoperative treatment methods be available to individuals that are suffering from this condition.

In July 2018, medical university researchers in Ireland lead a multi-national European research team to conclude in their research:

Platelet-rich plasma therapy is a simple, low-cost and minimally invasive intervention which is feasible to deliver in primary care to treat degenerative lesions of articular cartilage of the knee. This therapy appears to have minimal associated adverse events and may have beneficial effects in terms of pain, health utility, patient satisfaction and goal-orientated outcomes.(3)

What is interesting about this study is who the PRP helped:

Even a single injection of PRP provided benefit.

In the video below (there is no sound), I demonstrate how why administer PRP. In this case the patient has problems of meniscus degeneration. We apply multiply injections to support regeneration of the whole knee. Below the video is research that showed positive results of even a single PRP injection against a single placebo injection.

In November 2017, researchers reported on the benefits of PRP compared to placebo injection in patients who had osteoarthritis in both knees.Published in the American journal of physical medicine & rehabilitation, the study showedPRP treatment significantly improves pain, stiffness, and disability in patients with knee osteoarthritis compared to normal saline (placebo) treatment.(4)

Also in November 2017, in the International journal of rheumatic diseases, researchers reported a summary of the most recent findings on the benefits of PRP for knee osteoarthritis.

In the medical journal Arthroscopy, a journal devoted to obviously arthroscopy, surgeons are told thatPlatelet Rich Plasma injections(PRP),offers better symptomatic relief to patients with early knee degenerative changes (than hyaluronic acid or placebo), and its use should be considered in patients with kneeosteoarthritis.(6)

This is a verification of early research from the Mayo Clinic which came to the same conclusion PRPshowed better improvement than hyaluronic acid injection and placebo in reducing symptoms and improving function and quality of life. Especially in in younger, active patients with low-grade osteoarthritis.(7)

This is from the Mayo Clinic research:

Intraarticular platelet-rich plasma (PRP) injection has emerged as a promising treatment forkneeosteoarthritis. Studies to date, including multiple randomized controlled trials, have shown thatPRPis a safe and effective treatment option forkneeosteoarthritis. IntraarticularPRPis similar in efficacy to hyaluronic acid, and seems to be more effective than hyaluronic acid in younger, active patients with low-grade osteoarthritis. Treatment benefits seem to wane after 6-9 mos. There are numerousPRPtreatment variables that may be of importance, and the optimalPRPprotocol remains unclear.

At the end of that paragraph the Mayo team points out that benefits may only last 6 9 months and that there is much variation in the way PRP is delivered so it is difficult to understand why PRP may not work.

When we see a new patient with degenerative knee disease who had failed PRP treatments at other clinics, we ask them how many treatments did they have? More often than not they say one injection. For some with minor osteoarthritis, as pointed out by the medical studies highlighted in this article, one injection provides benefit.

But one injection may not be sufficient for someone who has a more active lifestyle than others.

Here is a recent study where the patients received two PRP injections as the complete PRP treatment program. This treatment group was considered to be active. This research was published in the journal Sports Health.(8)

A paper published in the Journal of physical therapy science.(9) It comes from doctors working in medical university hospitals in Turkey.

Highlights:

Below is what doctors are saying to each other about athletes who want to stay active. It was published in the medical journal Cartilage: As a result of the complexity of the arthritic knee, athletes, particularly those with a history of knee injury, have an earlier onset and higher prevalence of osteoarthritis. This can present a clinical dilemma to the physician managing the patient who, despite the presence of radiologically confirmed disease, has few symptoms and wishes to maintain an active lifestyle.(10)

The difficulty or challenge is in the prevention of advancing of knee osteoarthritis. Here the typical recommendations of anti-inflammatory medications, knee braces, and ice, those that the athlete can impose upon themselves, will lead to further knee deterioration. It is a challenge to convince an athlete of this when it may get them on the course, track of field this weekend.

In a recentstudy, researchers at Hospital for Special Surgery gavepatients with early osteoarthritis an injection of PRP (6-mL), and then monitored them for one year. At baseline and then one year after the PRP injection, physicians evaluated the knee cartilage with magnetic resonance imaging (MRI). While previous studies have shown that patients with osteoarthritis can lose roughly five percent of knee cartilage per year, the Hospital for Special Surgeryinvestigators found that a large majority of patients in their study had no further cartilage loss. At minimum PRP prevented further knee deterioration.

A leading provider of bone marrow derived stem cell therapy, Platelet Rich Plasma and Prolotherapy 11645 WILSHIRE BOULEVARD SUITE 120, LOS ANGELES, CA 90025

PHONE: (800) 300-9300

1 Huang Y, Liu X, Xu X, Liu J. Intra-articular injections of platelet-rich plasma, hyaluronic acid or corticosteroids for knee osteoarthritis. Der Orthopde. 2019 Jan 8:1-8. 2 Cook CS, Smith PA. Clinical Update: Why PRP Should Be Your First Choice for Injection Therapy inTreating Osteoarthritis of the Knee.Curr Rev Musculoskelet Med. 2018 Oct 22. doi: 10.1007/s12178-018-9524-x. 3 Glynn LG, Mustafa A, Casey M, et al. Platelet-rich plasma (PRP) therapy for knee arthritis: a feasibility study in primary care.Pilot Feasibility Stud. 2018;4:93. Published 2018 Jul 4. doi:10.1186/s40814-018-0288-2 4 Wu YT, Hsu KC, Li TY, Chang CK, Chen LC. Effects of platelet-rich plasma on pain and muscle strength in patients with knee osteoarthritis. American journal of physical medicine & rehabilitation. 2017 Nov. 5 Xing D, Wang B, Zhang W, Yang Z, Hou Y, Chen Y, Lin J. Intraarticular plateletrich plasma injections for knee osteoarthritis: An overview of systematic reviews and risk of bias considerations. International journal of rheumatic diseases. 2017 Dec 5. 6 Campbell KA, Saltzman BM, Mascarenhas R, Khair MM, Verma NN, Bach BR Jr, Cole BJ.A Systematic Review of Overlapping Meta-analyses.Arthroscopy. 2015 Nov;31(11):2213-21. doi: 10.1016/j.arthro.2015.03.041. Epub 2015 May 29. 7 Pourcho AM, Smith J, Wisniewski SJ, Sellon JL.Intraarticular platelet-rich plasma injection in the treatment of knee osteoarthritis: review and recommendations. Am J Phys Med Rehabil. 2014 Nov;93(11 Suppl 3):S108-21. doi: 10.1097/PHM.0000000000000115. 8 Gobbi A, Karnatzikos G, Mahajan V, Malchira S. Platelet-rich plasma treatment in symptomatic patients with knee osteoarthritis: preliminary results in a group of active patients.Sports Health. 2012;4(2):162-72. 9 Kavadar G, Demircioglu DT, Celik MY, Emre TY.Effectiveness of platelet-rich plasma in the treatment of moderate knee osteoarthritis: a randomized prospective study.J Phys Ther Sci. 2015 Dec;27(12):3863-7. doi: 10.1589/jpts.27.3863. Epub 2015 Dec 28. 10 Kirkendall DT. Management of the Retired Athlete with Osteoarthritis of the Knee. Cartilage January 2012 vol. 3 no. 1 suppl 69S-76S 11 Wang-Saegusa A, Cugat R, Ares O, et al. Infiltration of plasma rich in growth factors for osteoarthritis of the knee short-term effects on function and quality of life. Arch Orthop Trauma Surg. 2011 Mar;131(3):311-7. Epub 2010 Aug 17. 12 Injection of platelet-rich plasma in patients with primary and secondary knee osteoarthritis: a pilot study.Sampson S, Reed M, Silvers H, ey al. Injection of platelet-rich plasma in patients with primary and secondary knee osteoarthritis: a pilot study.Am J Phys Med Rehabil. 2010 Dec;89(12):961-9.1537-2290

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How Revolutionary Platelet-Rich Plasma Injections For Dogs …

By Platelet Rich Plasma Injections

This revolutionary treatment, which accelerates all kinds of healing, is now a favorite among vets for treating our beloved canine friends. It not just heals, but in many cases it revolutionizes how older dogs live their life. If youre an advocate for less invasive, safer, often less costly but effective, alternative treatment options, Platelet-Rich Plasma Injections are one of the rare kinds of treatment that fits the bill.

Treating dogs suffering with injuries to muscle, bone, or tendons has been by using Platelet Rich Plasma (PRP) injections has been shown to have a beneficial effect on recovery times, and the quality of the recovery.

Studies have shown that PRP promotes:

Two studies, one published in 2011 and 2013 by the US National Library of Medicine (NCIB) and the Journal of Surgical Research (JSR) respectively, demonstrate the beneficial therapeutic effects of PRP when administered to dogs with musculoskeletal injuries.

The 2011 study set out to report on the concentration of blood cells and growth factors in the animals used in the study. They sampled the blood cell density in untreated dogs and compared it to that of dogs that had received PRP. They also compared a number of growth factors between dogs that had been given PRP treatments and those that had not. They found that platelet counts in the treated animals were higher than they were in non-treated animals. Not all growth factors were found to have increased. Two sets of growth factors remained the same in the test and control populations. Despite the lack of change in a few growth factors, others showed a significant increase and the overall improvement to the therapeutic outcome had improved for the test animal population.

In the 2013 study, similar methods were used except two extra test groups were added; a control group where saline was added to the PHP solution, and a fourth group in which a random distribution of young and old beagles were tested. The results of this study brought the researchers to conclude that PRP alters the expression of certain targeted genes during the early stages of graft remodeling and that the platelet rich plasma can promote reinnervation and revascularization. They believe this explains the beneficial enhancement effect of PRP on graft maturation.

A study by Jeff Mayo DVM, Veterinary Medical Director of Ingeneron Inc. reports that his organization has been running studies on the effects of PRP on dogs. He says their studies have shown that they have proven that PRP is a safe and natural method to relieve the pain of injury or joint disease, promote healing and accelerate recovery time using the animals own blood. As a result of these findings, the Mobile Veterinary Surgical Services Serving of Seattle, WA are now confidently offering PRP treatments to the public in cases where the treatment is indicated.

The 2011 study mentioned above has been cited as conclusive evidence that that inflammation is reduced by PRP injection treatments and that enhanced healing results.

The KIM group (Kindred-Canines in Motion) reported in 2012 that PRP injections are safe and effective for most animals. KIM is now actively scheduling evaluations for possible candidates for PRP treatment of;

Further studies have found that PRP can be combined with Cold Laser therapies for even better cellular regeneration. In fact, they are complementary and synergistic. A research project published in Lasers Medical Science concluded that the deposition of collagen type I (a major contributing factor in regeneration) was higher when treatment with PRP and a Class 4 Cold Laser were combined, promoting an even faster rate of regeneration in the affected tissues.

And it looks like not all veterinary doctors are trained and equipped to use PRP treatments and Cold Laser therapy. As a result, owners who want their dogs to receive these treatments must go great lengths to seek out experienced practitioners. Fortunately, the awareness of these leading-edge treatments are spreading fast. Vets like Dr. R. Kraemer, who practices in orange county California and runs Vet4Bulldog.com, has expanded his offer for this groundbreaking combination to pet owners from anywhere in the State of California.

This combination works exceptionally well for joints and ligament healing which is a boon for older dogs. They are particularly affected by osteoarthritis and hence are badly in need for a combination treatment like this.

How laser therapy works Cold laser therapy is a non-invasive treatment that uses coherent light to induce cellular regeneration and improve circulation. The infrared laser light interacts with tissues at the cellular level.

Benefits of the combination treatment Cold Laser treatment increases cellular energy (ATP) to trigger an increase in cellular function, and health. Its light based and when it comes in contact with the cell, metabolic activity increases. Hence the improvement in the mobility of nutrients across cell membranes. Which means your pet gets healthier faster than ever.

Is PRP and Laser Combination Right for Your Dog? First of all, both PRP and Laser are noninvasive procedures. One uses growth factors in blood and the other uses light to stimulate cell regeneration and increase blood circulation. There isnt any other perfect combination of treatments that I can think of.

Almost Zero Downtime Both treatments may leave with soreness or inflammation, the typical side-effects of any injection/laser treatments. Under normal circumstances they are minor and will go away without any intervention. However, if symptoms are intense, icing the treatment area for 15 minutes 2-3 times a day for the first few days will help ease the pain and inflammation. Elderly animals and those with unusual conditions may require longer recovery times and closer attention.

Vets whove tried these report, that in most cases the results are almost instantaneous (as in obvious from day 1.) And as high as 80% of the animal patients treated with PRP have a noticeable improvement in their mobility within fewer than 10 days. Pets can resume normal activity in under a week: but may need recovery and rehabilitation therapies tailored to its condition, age, temperament, goals, and abilities.

As far as precautions are concerned, there isnt any to worry about. Except that for PRP, the animal patient needs to be off Anti-inflammatory medication at least a week prior to the treatment. Other than that, PRP comes with very few side effects. The same is the case for cold Laser therapy: zero known side effects. That is other than the obvious contraindications. Animals with any of the following conditions should not receive Cold Laser treatment.

PRP improves the quality of a dogs life as well as the life of its owner. Older dogs and dogs with osteoarthritis can benefit tremendously from PRP as it is a low-intensity treatment.

With emerging knowledge of the positive effects on the health and well-being PRP and Cold Laser treatments have on our animals, the only things standing between many dogs and better health is a lack of public awareness and the availability of these treatments.

Although these treatments look simple and easy to the untrained eye, theres more to it than what meets the eye. Experience counts. Proper training is a requirement. You dont want your vet to practice these treatments on your pet. So do your due diligence. Ask your vet if he/she does these treatments on a regular basis.

One of the things you need to clearly verify with your vet is the number of treatments required. Depending on the condition of your pet, your vet may prescribe 3-5 treatments. And thats perfectly okay. Because weve seen that repeated treatments often produce better results.

Also ask if theyll use sedatives.

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Top Stem Cell Conferences | Stem Cell Research 2019 …

By Stem Cell Medicine

Session and Tracks Track 1: Stem Cells Biology

Stem cells are defined as precursor cells that have the capability to self-renew and to come up with multiple mature cell types. Stem cells are an ongoing source of the differentiated cells that make up the tissues and organs of plants and animals. After collecting and culturing tissues is it possible to classify cells as per their operational concept. There are 2 major types of stem cells: Embryonic Stem Cells and Adult Stem Cells that is also known as tissue stem cells. This difficulty in characteristic stem cells in situ, without any manipulation, limits the understanding of their true nature. There is great interest in stem cells as a result of they have potential in the development of therapies for replacing defective or damaged cells ensuing from a variety of disorders and injuries, like Parkinson disease, heart disease, and diabetes.

Related Conferences:

25thGlobal Meet on Cancer Research & Oncology, May 20-21, 2019, Rome, Italy; 2ndWorld Congress on Advanced Cancer Science & Therapy, January 28-29, 2019, Dubai, UAE; 3rdAdvances in Cell & Stem Cell Research Congress, September 25-26, 2019, Rome, Italy; 3rdInternational Conference on Nanostructures, Nanomaterials and Nanoengineering, October 21-22, 2019, Las Vegas, USA; 3rdWorld Congress on Advanced Biomaterials and Tissue Engineering, August 26-27, 2019, Madrid, Spain;

Hematopoietic Stem Cells are the immature cell that is developed into all types of blood cells, including red blood cells, white blood cells, and platelets which are found in the peripheral blood and the bone marrow. These stem cells are also called blood stem cell. Studies have described two populations of Hematopoietic Stem Cells that are Long Term and Short Term. Long-Term Hematopoietic stem cells which are capable of self-renewal, while Short Term Hematopoietic stem cells do not have this capacity.

Related Conferences:

4thWorld Biotechnology Congress, May 20-21, 2019, London, UK; 6thWorld Congress on Microbial Biotechnology, June 17-18, 2019, Paris, France; Annual Congress on Advanced Tissue Science and Regenerative Medicine, April 15-16, 2019, Amsterdam, Netherlands; World Congress on Cell & Gene Therapy, September 25-26, 2019, Rome, Italy; World Congress on Novel Trends and Advances in Biotechnology, September 25-26, 2019, Rome, Italy;

Embryonic Stem Cells are developed when embryos formed during the blastocyst phase of embryological development. They can grow in all derivatives of the 3 primary germ layers i.e. ectoderm, endoderm and mesoderm. These include each of the more than 220 cell varieties within the adult body. Pluripotency distinguishes embryonic stem cells from adult stem cells found in adults; whereas embryonic stem cells can generate all cell types within the body, adult stem cells are multipotent and can produce only a restricted number of cell types. Embryonic stem cells are capable of propagating themselves indefinitely. This allows embryonic stem cells to be employed as useful tools for both research and regenerative medicine, because they can produce limitless numbers of themselves for continued research or clinical use.

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Induced Pluripotent Stem Cells (iPSCs) are the adult stem cells derived from skin or blood cells which are reprogrammed to an embryonic stem cell maintaining the essential properties of introducing important genes and also to enables the development of an unlimited source of any type of human cell needed for the therapeutic purpose. Researchers have rapidly developed the techniques for generating iPSCs and by creating a new and powerful way to "de-differentiate" cells whose developmental fates.

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Though the concept of stem cell niche was prevailing in vertebrates, the first characterization of stem cell niche in vivo was figured out in drosophila germinal development. A stem-cell niche is an area of a tissue that provides a specific microenvironment, in which stem cells are present in an undifferentiated and self-renewable state. Cells of the stem-cell niche interact with the stem cells to take care of them or promote their differentiation. Characterization of these stem cell niches depends on the ability to identify stem cells in vivo in their normal setting. Through comparison of different stem cell systems, some themes emerge that indicate possible general characteristics of the relationship between stem cells and their supporting niche.

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Stem cell banking is the extraction, processing and storage of stem cells which can used for treatment when required. Stem cells have the amazing power to transform into any tissue or organ in the body. It is due to this unique characteristic that they have the potential to treat over 80 life threatening diseases, and provide numerous benefits to the baby, its siblings and the family. There are variety of sources from where stem cells can be banked, with the most common amongst them being the umbilical cord. Cord blood banking is that the extraction of stem cells from the umbilical cord. This is done during childbirth and is a fast, hassle free and painless procedure. While, the umbilical cord and cord blood are the foremost common sources of stem cells - the Placenta, amniotic sac and amniotic fluid are by far the richest sources, in terms of both - quantity and quality. Some other rich sources of stem cells are Placenta, Umbilical Cord, Amniotic Fluid, Dental Stem Cells, Menstrual Fluid, Adipose Tissue and Bone Marrow.

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A procedure in which a patient receives healthy blood-forming cells (stem cells) to replace their own, when stem cells or bone marrow are damaged or destroyed by disease, as well as some types of cancer, or by high doses of chemotherapy or radiation therapy used to treat cancer. The healthy stem cells may come from the blood or from a donors bone marrow or from the umbilical cord blood of a newborn baby. A stem cell transplant may be autologous (use of stem cells from your own bone marrow or blood), allogeneic use of stem cells from someone else, the donor could also be a relative or somebody who isn't associated with you) or syngeneic (use of stem cells from an identical twin). The stem cells within the bone marrow transform into red blood cells, white blood cells and platelets. when these blood cells mature, they go into the peripheral blood (the blood that flows through the body). If the bone marrow is damaged or destroyed, it cant create normal blood cells. in a stem cell transplant, healthy stem cells are placed in your body to assist your bone marrow start to work properly. The new stem cells make healthy blood cells.

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Stem Cell Therapy is the treatment for various disorders which non-seriously life-threatening by using stem cells. These stem cells can be obtained from a lot of different sources and used to potentially treat more than 80 disorders which include neuromuscular, organ, chronic and degenerative disorders. Chronic disorders arise from degeneration or wear and tear of cartilage, muscle, bone, fat or the opposite organ, tissue or cell. This may occur owing to a spread of reasons, but it's usually the tactic spoken as aging, or 'getting old' that is the largest cause. Stem cell therapy is currently being researched for the treatment of various diseases. While research and clinical trials are in process with varying degrees of success, stem cell therapy holds the potential to offer a successful cure for these conditions.

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Cancer is defined as the abnormal growth of cells that possesses the ability to spread to other cells and tissues. Cancer is one of the major illness which it seemed to be more prevalent all over the world. Even though the death rate and peoples suffering from these diseases are in greater number in recent years. There are over 200 variety of types of cancer across the globe. The death rate increasing year-by-year due to this disease even in developed countries. Cancer Stem Cells (CSCs) are a small population of cells inside tumors with capabilities of self-renewal, differentiation, and tumorigenicity once transplanted into an animal host. The CSC hypothesis thus doesn't imply that cancer is always caused by stem cells or that the potential application of stem cells to treat conditions like cardiovascular disease or diabetes which is able to result in tumor formation. Rather, tumor-initiating cells possess stem-like characteristics to a degree sufficient to warrant the comparison with stem cells along with the observed experimental and clinical behaviors of metastatic cancer cells are extremely resembling the classical properties of stem cells

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As an organism grows and develops, carefully orchestrated chemical reactions activate and deactivate components of the genome at strategic times and in specific locations. Epigenetics is that the study of these chemical reactions and the factors that influence them. It is strongly believed that there are some signals at the epigenetic level that regulate the fate of the stem cells. Though all of the cells in our body contain the same genetic makeup. These genes are not necessarily active at all times, rather they are expressed at times when needed in a highly controlled fashion.

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Tissue Engineering is a scientific field centered on the advance of Tissue and Organ Substitutes by controlling their environment, biomechanical and biophysical parameters which include the utilization of a different or same scaffold for the arrangement of new tissue. These frameworks empower the In-vitro investigation of human physiology and physiopathology, while giving a rendezvous of biomedical instruments with potential materialness in toxicology, medicinal gadgets, tissue substitution, repair and Regenerative Medicine. Regeneration is that the progression of renewal, regeneration, and growth that makes it cells, organ regeneration to natural changes or events that cause damage or disturbance. This study is carried out as craniofacial tissue engineering, in-situ tissue regeneration, adipose-derived stem cells for tissue science which is also a breakthrough in cell culture technology. The study isn't stopped with the regeneration of tissue wherever it is further carried out in relation to cell signaling, morphogenetic proteins. Most of the neurological disorders occurred accidentally having a scope of recovery by replacement or repair of intervertebral discs repair, spinal fusion and plenty of more advancement.

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Regeneration Medicine is that the Survival of any living body essentially relying upon its capability to repair and recover injured or harmed tissues or potential organs amid its lifespan following injury, illness, or maturing. This will shape the system for recognizing novel clinical medicines which will enhance the mending and regenerative limit of individuals. The Regeneration process involves Cell Proliferation where most of the medical disorders occurred accidentally includes a scope of recovery by replacement or repair of intervertebral discs repair, spinal fusion and plenty of more advancements.

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Cell reprogramming is the process of reverting mature, specialized cells into induced pluripotent stem cells. Reprogramming also refers to the erasure and re-establishment of epigenetic marks during mammalian germ cell development. The discovery of Induced pluripotent stem cells emphasizes on reprograming of any adult differentiated cells into stem cells by genetic modification under precisely controlled laboratory conditions. Reprograming of cells is supposed to presage revolution in both, medical and biological research and allows modeling and analysis of human diseases and cell cytotoxicity by drugs. The technique is still in its growing phase and requires a great deal of extensive research and approval from authorities for further trials.

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Stem cell nanotechnology has emerged as a brand-new exciting field. Experimental and theoretical studies of interaction between nanostructures or nanomaterials and stem cells have made great advances. The importance of nanomaterials, nanostructures, and nanotechnology to the basic developments in stem cells-based therapies for injuries and degenerative diseases has been recognized. Apart from tracking the localization of stem cells, nanotechnology has improved targetability, half-life, and stability of stem cells by providing a suitable microenvironment. In particular nanomaterials have played a significant role in the isolation and proliferation or differentiation of stem cells and intracellular delivery of small and macromolecules within stem cells. In this field over the past few years, explore the application prospects, and discuss the issues, approaches and challenges, with the aim of rising application of nanotechnology in the stem cells research and development.

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Stem Cell Technologies will become a global biotechnology that manufactures, develops and sells product by providing the services to support academic and industrial scientists. Stem cells analysis and development team typically collaborates with educational institutes and industrial partners to manufacture, develop and distribute a specific product for a given analysis. A stem cell has helped several scientific communities and industries to develop technologies to achieving the world biotechnology market. The corporate makes a specialist in developing cell culture media, cell separation product, instruments and completely different reagents to be utilized in the cell, immunology, cancer, Regenerative medicine and cellular treatment analysis.

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There are many research advancements and applications and of Stem Cells. Stem cell research that can be applied to develop new therapies includes cell replacement therapy, development of drugs, using iPSC technology to generate stem cells from the patients skin or blood, using trans differentiation technology to convert a specialized cell type to a progenitor cell and many more. It also carries the immense potential for treating a number of human diseases such as to repair or regenerate blood vessels, treatment of eyesight, Diabetes, Neurodegenerative Disorders and Wound Healing etc.

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Carefully planned and ethically approved clinical trials resulting from a robust preclinical pathway are necessary to advance the field. This will require a programmatic approach that involves partnerships of clinicians, academics, industry, and regulatory authorities with a focus on understanding basic biology that informs a tight linkage between preclinical and clinical studies. Rather than suggesting that clinical trials are premature, such trials should be encouraged as part of multidisciplinary programs in regenerative medicine.

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The field of bioethics has addressed a broad swathe of human inquiry, ranging from debates over the boundaries of life, surrogacy, and the allocation of scarce health care resources to the right to refuse medical care for religious or cultural reasons. StemGen is a research database of international, regional and national normative instruments concerning the socio-ethical and legal aspects of stem cell research and related therapies. The regulation of stem cell research is an issue that has drawn much comment, criticism and even judicial arbitration in recent years along with the marketing status of Stem Cells, Cell therapy, Regenerative Medicine, Tissue Engineering and many more worldwide.

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Differentiation of human induced pluripotent stem cells into …

By Induced Pluripotent Stem Cells

Culture of human iPSCs

The commercial human iPSCs used in this study were purchased from Saibei Biotechnology (Beijing, China). iPSCs (HiPSC-U1) were reprogrammed from human urine-derived cells of a 37-year-old male by the integration-free CytoTune-iPS 2.0 Sendai Reprogramming Kit (Thermo Fisher Scientific, MA, USA). iPSCs were cultured in 1% Matrigel-coated (BD Biosciences Co., Ltd., NM, USA) Petri dishes with E8 medium (Saibei Biotechnology) at 37C and 5% CO2. The medium was refreshed daily. iPSCs were passaged once every 6 days with 0.25% ethylenediaminetetraacetic acid (EDTA, Saibei Biotechnology). 1mL of 0.25% EDTA was added and the cells were placed at 37C and 5% CO2 for 5min. When iPSC colonies appeared white, the solution was gently removed, and the iPSCs were washed with Mg2+ and Ca2+-free Dulbeccos PBS (Sigma, MO, USA). iPSCs were then harvested by gently pipetting 710 times with 1mL E8 medium and seeded onto fresh six-well culture plates that were coated with 1% Matrigel at a ratio of 1:6. 10m Y-27632 (Sigma) was supplemented in the medium on the first day of passage.

Human LCs were obtained from nine male donors with a mean age of 45 years old through testes excision within 20h. Informed consent was obtained from each donor, and this study was approved by Human Research and Ethical Committee of Wenzhou Medical University. The testes were used to isolate ILCs. ILCs express all androgen biosynthetic enzymes56, and are capable of proliferation and differentiation57. The isolation of ILCs was performed as previously described56. In brief, the testes were perfused with collagenase (Sigma) via the testicular artery, and digested with M-199 buffer (Gibco, NY, USA) containing collagenase (0.25mg/ml) and DNase (0.25mg/mL, Sigma) for 15min. Then, the cell suspension was filtered through 100m nylon mesh and the cells were separated by a Percoll gradient (Sigma). The cells with the density of 1.071.088g/ml were collected. The purity of ILCs was evaluated by immunohistochemical staining HSD3B1, the biomarker of ILCs, as previously described58. The HSD3B1 staining solution contained with 0.4mm etiocholanolone (Sigma) as the steroid substrate and NAD+ as a cofactor58. The purity of ILCs was >95%.

The isolated ILCs were directly seeded into wells in the 24-well culture plates with the density of 2104 cells/well and incubated at a 37C, 5% CO2 incubator. The culture medium (LC-Medium) contains DMEM/F12 (Gibco), 5% fatal bovine serum (FBS, Gibco), 2.5% horse serum (HS, Gibco), and 1% penicillin/streptomycin (P/S, Gibco). In order to get ALCs, the culture medium were changed into differentiation-induced medium (DIM) contains DMEM/F12, 5mm ITS (insulin, transferrin, and selenium, Sigma), 5ng/ml luteinizing hormone (LH, PeproTech, NJ, USA), and 5mm lithium chloride (Li, Sigma) as our team previous report35.

The Sprague-Dawley rats (at 5 weeks of age) and immune deficiency (SCID) mice (at 5 weeks of age) were obtained from the laboratory animal center of Wenzhou Medical University, Wenzhou, China. All animals were kept under conditions with controlled temperature (232C), a 12h dark/light cycle, and relative humidity of 4555%. The standard drinking water and rodent diet were accessed ad libitum. All surgical procedures and postoperative care were approved by the Wenzhou Medical Universitys Animal Care and Use Committee, and were performed in accordance with the Guide for the Care and Use of Laboratory Animals.

The point at which iPSCs were expanded to ~70% confluency in the E8 medium was defined as day 2, and at this point when iPSCs were changed into E7 medium (no FGF2) for 2 days to prepare differentiation. From day 5 to 0, the medium was refreshed daily. Prior to the beginning of differentiation, iPSCs were cultured in a differentiation-inducing medium composed of DMEM/F12, 1% bovine serum albumin (BSA) (Sigma), 5mm ITS, 5ng/mL LH. From 07 days, 0.2m SAG (DHH agonist, Sigma), 5m 22R-OHC (Steraloids, RI, USA), and 5mm Li were added into iPSC-DIM. From 710 days, 5ng/mL PDGF-AA (Sigma) and 5ng/mL FGF2 (Sigma) were added into iPSC-DIM. From 1017 days, 5ng/mL PDGF-AA, 5nM IGF1 (Sigma), and 10m Androgen (Sigma) were added into iPSC-DIM. From 1720 days, 10ng/mL PDGF-AA and 10ng/mL FGF2 were added into iPSC-DIM. From 2025 days, 5ng/mL LH, 0.5mm retinoic acid (RA, Sigma) and 1mm 8-Br-cAMP (Sigma) were added into iPSC-DIM. From day 0 to 25, the medium was changed every 2 days by fresh iPSC-DIM. From 2530 days, the cells were mechanically enriched by scraping away clonal iPSC-like cells. The remaining Leydig-like cells were kept in Enrichment Medium contained DMEM/F12, 5% FBS, 2.5% HS, 1sodiumpyruvate (Invitrogen), 1GlutaMAX (Invitrogen), and 1% P/S for the subsequent assays. The medium was changed every 2 days by fresh Enrichment Medium.

For TEM, the cells in different groups were prefixed with 2.5% glutaraldehyde in 0.1m PBS for 24h at 4C. Then, they were washed with PBS, and post-fixed with 1% osmium tetroxide. After gradient dehydration of acetone, they were embedded in Araldite M (Sigma Aldrich). Ultrathin sections (1m) were subsequently cut with an ultramicrotome, mounted on nickel grids, and stained with uranyl acetate and lead citrate. At last, the samples were sent to the electron microscope room at Wenzhou Medical University for subsequent processing and testing using a transmission electron microscope (H-600A-2; Hitachi, Tokyo, Japan).

The cell culture supernatants and serum were collected at each experimental time point for the quantitative measurement of testosterone. For the cell culture supernatants, 10ng/mL LH was in advance at least 3h added into the medium (just having DMEM/F12) to stimulate the testosterone production of LCs or iPSC-LCs. Testosterone levels were measured with a tritium-based radioimmunoassay using anti-testosterone antibody as previously described59. Standards ranging between 10 and 2000pg/mL testosterone were prepared in triplicate. Standards and samples were incubated with tracer and antibody at 4C overnight and charcoal-dextran suspension was used to separate the bound and free steroids. The bound steroids were mixed with a scintillation buffer and counted in a scintillation counter (PE, CA, USA). The minimum detectable concentration for testosterone was 5pg/mL. Quality control samples contain 100pg/mL testosterone. The intra-assay and inter-assay coefficients of variation were within 10%.

Immunofluorescence was used to identify iPSC-LCs as a previous report60. In brief, after fixation with 4% paraformaldehyde (Sigma) for 15min, cells were washed three times with PBS. Then cells were permeabilized with 0.1% TritonX-100 in PBS for 15min at room temperature, and incubated with 3% (w/v) BSA in PBS for 1h at room temperature. The cells were then incubated with primary antibodies as TableS1 overnight at 4C, and then with fluorescein isothiocyanate (FITC)-conjugated anti-mouse, FITC-conjugated anti-rabbit, Cy3-conjugated anti-mouse, and Cy3-conjugated anti-rabbit IgG secondary antibodies (1:1000, Bioword, USA) for 60min at room temperature. Then the cells were rinsed three times with PBS thrice for 5min each and then incubated for 15min with DAPI (Sigma) for nuclear staining and washed three times with PBS before examination by an inverted fluorescence microscope (OLYMPUS, Japan).

Total RNA from the cells was extracted using Trizol reagent (Invitrogen, CA, USA) according to the manufacturers instruction. The RNA was reversely transcribed into cDNA using the Superscript II kit (Invitrogen). The cDNAs templates were diluted 1:10, which were used to perform RT-PCR and qPCR to analyze the gene expressions. RT-PCR was performed using an authorized thermal cycler (Eppendorf, Hamburg, GER). After amplification, 1L of 6Loading buffer and 5L of each PCR product were mixed and electrophoresed on a 2% agarose containing 0.5g/mL ethidium bromide. Gels were scanned for further analysis. qPCR was performed using the Thunderbird SYBR qPCR Mix (Takara, Tokyo, Japan) according to the manufacturers instructions. Signals were detected using a Light Cycler 480 Detection System (Roche, Basel, Switzerland). The relative expression of genes was normalized to GAPDH. The melting curve was examined for the quality of PCR amplification for each sample, and quantification was performed using the comparative 2-Ct method. The primer sequences were shown in TableS2.

Total RNA from each sample was extracted using Trizol reagent (Invitrogen, CA, USA). 12g total RNA was used to prepare the sequencing library. To sequence the libraries, the barcoded libraries were mixed, denatured to single stranded DNA, captured on Illumina flow cell, amplified in situ, and subsequently sequenced for 150 cycles for both ends on Illumina HiSeq 4000 instrument. Sequence quality was examined using the FastQC software. The transcript abundances for each sample were estimated with StringTie, and the FPKM value for gene and transcript level were calculated with R package Ballgown. The differentially expressed genes and transcripts were filtered using R package Ballgown. The correlation analysis was based on gene expression levels. Hierarchical Clustering, Gene Ontology, Pathway analysis, scatter plots and volcano plots were performed with the differentially expressed genes in R, Python for statistical computing and graphics61.

Cells were washed with cold PBS and were lysed in 1radioimmunoprecipitation assay lysis buffer in the presence of a protease inhibitor mixture/1% phosphatase inhibitor mixture (Roche). 50g of protein samples were applied to a 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis and transferred into the polyvinylidene difluoride membranes (Sigma) by an electroblot apparatus. After being blocked with a blocking solution (5% fat-free milk) for 2h at 4C, the membranes were incubated with primary antibodies as TableS1 in the blocking solution at 4C overnight. The membranes were washed with tris-buffered saline with Tween 20 (TBST) five times (10min each), and incubated with horseradish peroxidase-conjugated secondary antibody (1:3000, Bioword) at room temperature for 2h. The membranes were then washed five times (10min each) with TBST. Bands were visualized with enhanced chemiluminescence (ECL, Pierce, USA). The protein expression was normalized to -actin.

The cell samples were fixed with 4% paraformaldehyde in PBS and permeabilized with 0.1% TritonX-100 (Sigma). The samples were then labeled with primary or isotype control antibodies for 30min at 4C. Primary and isotype control antibodies that were not conjugated to fluorophores were labeled with fluorophore-conjugated secondary antibody for 30min at 4C. The labeled samples were detected by flow cytometry analyzer (BD, USA). Data analysis was performed on FCS Express 4 Flow Research Edition software.

The standard protocol was conducted as PKH26 Product Information Sheet (Sigma, MINI2). In brief, the suspension containing 2107 cells were centrifuged (400g, 5min) and were washed once using fresh medium without serum. After centrifuging, the supernatant was removed and no more than 25L of 2Cell Suspension was prepared by adding 1mL of Diluent C, the cells were resuspended with gentle pipetting to ensure complete dispersion. 2Dye Solution (4106m) was prepared by adding 4L of the PKH26 ethanolic dye solution to 1mL of Diluent C and mixed well. Then 1mL of 2Dye Solution was rapidly added into the 1mL of 2Cell Suspension. Final concentration after mixing was 2106m PKH26 with 1107 cells/well. The mixing suspension was incubated at room temperature for 5min with periodic mixing. The action of the staining was stopped by adding an equal volume (2mL) of serum. Then the suspension was centrifuged at 400g for 10min and washed three times. Finally, the cells tagged with PKH26 were seeded on fresh wells and used for injection.

For evaluating whether iPSC-LCs could facilitate the recovery of LC dysfunction of rats, iPSC-LC transplantation was performed as previously described with some modifications62. Sixty 49-days-old male Sprague-Dawley rats (n=5 for each group at each time point) were used in this study. Before transplantation, male rats were administered a single intraperitoneal injection of EDS (75mg/kg, Pterosaur Biotech Co., Ltd., Hangzhou, China), which was dissolved in dimethyl sulfoxide (Sigma): H2O (1: 3, v/v). This treatment resulted in the elimination of LCs in the adult testes of rats63. Then iPSC-LCs labeled with PKH26 (red) were resuspended manually, and harvested in a 15mL Falcon tube. Cells were rinsed twice with PBS following centrifugation at 200g for 5min. Finally each pellet was resuspended in PBS for transplantation. Cells were loaded into a 1mL syringe for injection into the testis of adult Sprague-Dawley male rats that had been treated with EDS. Approximately 2106 PKH26-labeled iPSC-LCs in 40mL of PBS were injected into the parenchyma of recipient testes 7 days after the rats received EDS. The control animals for the experimental group were EDS-treated rats that had received a testicular injection of the PBS vehicle. Testes from all animals were examined at 0, 7, 14, and 21 days after EDS treatment.

One testis from each rat was used for immunohistochemistry (Vector Laboratories, Inc., Burlingame, CA, USA) according to the manufacturers instructions. The rats were killed with an overdose of sodium pentobarbital (Sigma). Testes were removed and fixed in 4% paraformaldehyde overnight at 4C. Then testes were dehydrated with a graded series of ethanol and xylene and subsequently embedded in paraffin. Five micrometer-thick transverse sections (5m) were cut, de-waxed in water, and were mounted on glass slides. Antigen retrieval was performed by microwave irradiation for 10min in 10mm (pH 6.0) of citrate buffer, after which endogenous peroxidase was blocked with 0.5% of H2O2 in methanol for 30min. Some sections were fixed with 4% paraformaldehyde for 15min and washed 3 times with PBS. Then they were permeabilized with 0.1% TritonX-100 in PBS for 15min at room temperature, and incubated with 3% (w/v) BSA (Sigma) in PBS for 1h at room temperature. Then these sections were then incubated with an CYP11A1 polyclonal antibody diluted 1:1000 for 2h at room temperature, and then with FITC-conjugated IgG secondary antibodies (1:1000, Bioword) for 1h at room temperature. These sections were rinsed with PBS three times for 5min each time. Then the sections were incubated for 15min with DAPI (10g/mL, Sigma) for nuclear staining and washed three times with PBS. The sections were cover-slipped with resin (Thermo Fisher Scientific, Waltham, UK). At last, they were examined by an inverted fluorescence microscope (OLYMPUS). The cells with CYP11A1 staining in the interstitial area represent the LC64.

Other sections were directly incubated with CYP11A1 polyclonal antibody diluted 1:1000, for 2h at room temperature. Diaminobenzidine was used for visualizing the antibodyantigen complexes, positive labeling LCs by brown cytoplasmic staining. Mayer hematoxylin was applied in counterstaining. The sections were then dehydrated in graded concentrations of alcohol and cover-slipped with resin (Thermo Fisher Scientific, Waltham, UK). Lastly, they were examined by a fluorescence microscope (LEICA). The cells with CYP11A1 staining in the interstitial area represent the LCs.

For teratoma formation, iPSCs (5106 cells) were dissociated with 0.5mm EDTA, centrifuged, resuspended in 100L E8 with 1% Matrigel, and injected into the hind limbs of 6-week-old male SCID mice. Teratomas were collected after 6 weeks, and fixed in 4% paraformaldehyde for paraffin embedding and hematoxylin and eosin staining. Slides were imaged and analyzed by a qualified clinical pathologist.

The division of iPSCs was blocked with 50g/mL of colcemid solution (Invitrogen, USA). Cells were washed with PBS and harvested with trypsin at room temperature for 2min. Then cells were fixed in methanol/glacial acetic acid (3:1) for three times and dropped onto slides for chromosome spreads. At last, the slides were baked at 55C for overnight. Standard G-banded karyotypes were obtained using Giemsa solution staining (Giemsa, Japan).

To enumerate CYP11A1-positive Leydig cell numbers, sampling of the testis was performed according to a fractionator method as our previous report65. Identification of all Leydig cell lineages was done by the staining of CYP11A1. About 10 testis sections per rat were sampled from each testis. The total number of LCs was calculated by multiplying the number of LCs counted in a known fraction of the testis by the inverse of the sampling probability.

All experiments were performed at least thrice, and the data are presented as the meanstandard error of the mean. Statistical analyses were evaluated using an unpaired Students t test or one-way analysis of variance for more than two groups. P<0.05 was considered statistically significant.

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Differentiation of human induced pluripotent stem cells into ...

Cardiovascular Division at Miller School of Medicine

By Uncategorized

Jeffrey J. Goldberger, M.D., M.B.A., has been named Chief of the Cardiovascular Division at the University of Miami Miller School of Medicine, adding to the Divisions already stellar faculty. He will also oversee all cardiac clinical care at UHealth the University of Miami Health System. A renowned electrophysiologist who has published nearly 200 studies, Goldberger joins the Miller School this week from Northwestern University Feinberg School of Medicine, where he was professor of medicine and Director of the Program in Cardiac Arrhythmias in the Center for Cardiovascular Innovation.

We are extremely invigorated to have Dr. Goldberger joining the Miller School of Medicine, bringing his commitment to breakthrough research and innovative clinical care to South Florida, said Roy E. Weiss, M.D., Ph.D., professor and Chairman of Medicine at the Miller School. He shares our commitment to translating research into personalized clinical care.

Goldberger plans to expand the multidisciplinary team approach to detecting, diagnosing, and successfully treating atrial fibrillation, developing novel ablation procedures to treat arrhythmias in patients at UHealth.

With the excellent clinical structure UHealth has established in the region, there is a rich environment for innovation and discovery, said Goldberger, who will be professor of medicine at the Miller School. He will also integrate cardiology services performed by UHealth doctors at Jackson Memorial Hospital and the Miami VA Healthcare System.

We have a unique opportunity to leverage our expertise in electrophysiology, interventional cardiology, advanced heart failure, vascular treatments and imaging to offer patients multidisciplinary therapies, Goldberger said. I hope to accelerate the pace of discoveries and enhance our translational research to open new opportunities to bring the discoveries to the patients who need them.

With continuous funding from the National Institutes of Health and the American Heart Association over the last 12 years, Goldberger has a history of leading novel trials that shift the standard of care. Most recently, he led the OBTAIN (Outcomes of Beta-Blocker Therapy After Myocardial Infarction) study, which found that heart attack patients treated with a substantially lower dosage of beta-blockers than used in earlier trials survived at the same rate, or even better, than those receiving higher doses. The study was published in the September 21, 2015, Journal of the American College of Cardiology.

Goldberger leads a national think tank on sudden cardiac death, continuing the decade-long work he has done with Robert Myerburg, M.D., professor of medicine at the Miller School and a renowned expert in the field. After receiving his medical degree and completing his residency at Albert Einstein College of Medicine, Goldberger completed a fellowship in cardiology at the University of California, San Francisco. He has served as an editorial consultant on nearly two dozen professional or peer-reviewed journals, including the Journal of the American Medical Association, Circulation and the New England Journal of Medicine.

Goldbergers wife, Sharon Sholiton, M.D., is Associate Dean for Student Affairs at Rush University Medical School in Chicago, and they have four children.

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Cardiovascular Division at Miller School of Medicine

Mayo Clinic Q and A: Stem cell therapy for arthritis …

By Stem Cell Clinic

DEAR MAYO CLINIC: Whats the latest information on using stem cell therapy to treat an arthritic shoulder that causes excessive pain?

ANSWER: New efforts in regenerative medicine, including stem cell therapy, could dramatically affect orthopedic surgery over the coming years. Much of this hope is pinned on using stem cells to treat degenerative conditions such as shoulder arthritis. Although it shows promise, stem cell treatment for arthritis isnt widely available at this time, as its still being researched.

Stem cells are the basic building blocks of all human tissue. Stem cells hold potential as treatment, in part, because they can communicate valuable information about tissue growth and healing to other cells in the body. Arthritis involves joint degeneration due to loss of the cartilage that cushions bones. Recently researchers have begun to look to stem cells for orthopedic conditions such as shoulder arthritis. Progress using stem cells to treat arthritis already has been reported, with the ultimate goal of using stem cells to regrow cartilage.

When discussing stem cell therapy, its important to understand that pure stem cells are not currently available to U.S. patients outside of a clinical research study. A handful of clinical research trials, monitored by the U.S. Food and Drug Administration (FDA), are ongoing at this time to study stem cell treatment for arthritis. The early findings from these trials are encouraging.

Unfortunately, the excitement surrounding emerging stem cell therapy has led some patients and health care providers to overlook the lack of scientific evidence to support its use at this time. Stem cell therapies currently used outside clinical studies do not contain pure stem cells. Instead, they are a mix of a variety of cells, of which only a very small percentage are stem cells. It is possible that many of these treatments do not contain enough stem cells to help.

It is also important to recognize that many stem cell therapies now marketed directly to patients are conducted without the required biologics license from the FDA. Also, some forms of mislabeled stem cell therapies do not contain any living stem cells. Such practices are cause for concern, as these treatments can mislead patients and the public, and delay the scientific progress needed to turn stem cell therapies into cures.

What the research into stem cells and arthritis shows is that there are opportunities for stem cell treatment to be used as injection therapy alone and in addition to orthopedic surgical procedures. Successful stem cell therapies thus far have resulted mostly in pain relief and improvement in function or quality of life. Only a few limited early studies have demonstrated improvement in new cartilage or bone formation needed to cure arthritis. Exactly how that cartilage regrowth occurs, or even how pain relief is achieved, is still unknown. That means if you have a stem cell procedure, it will be used to treat the symptoms of arthritis only. The ability to cure the disease entirely is not yet available.

No major research studies have specifically investigated stem cell treatment for shoulder arthritis. Much of what is known about stem cells in arthritis comes from research into knee degeneration. Its not known if the successes treating knee arthritis will prove to be similarly beneficial when used for the shoulder. Therefore, current recommendations to treat shoulder arthritis remain the judicious use of gentle pain relievers, exercise and occasional steroid injections. In severe cases, shoulder replacement can provide long-lasting pain relief.

With demonstrable safety and mounting evidence of the effectiveness of stem cell therapy for some orthopedic conditions, potentially all orthopedic disease could be treated with stem cell therapy in the future. But, first, doctors and patients will have to wait until the scientific evidence catches up to the excitement around this promising option. Dr. Shane Shapiro, Orthopedic Surgery and Center for Regenerative Medicine, Mayo Clinic, Jacksonville, Florida

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Mayo Clinic Q and A: Stem cell therapy for arthritis ...

COPD and stem cell treatment | Mayo Clinic Connect

By Stem Cell Clinic

I have been trying to find some actual people who have tried any of the Stem Cell treatments being offered. Especially interested in the Cellular Therapy being offered at many clinics in the USA. Not knowing who or what to believe is very difficult. I have also heard that Aloe Vera is an effective treatment that can help immensely I don't know how much of this is true either. I am on oxygen therapy 24/7 and my nose runs so much that I am desperate. I have tried Saline Solution, Sesame seed oil, Aloe Vera from the plant inside my nose. I have a humidity container on my oxygen consentrator and also a humidifier in my bedroom. I have been trying to take off the oxygen for an hour or so every few hours but my oxygen gets low and I go into A-fib. Nothing has worked to stop my runny nose. It truly makes me want to just give up. I am also in remission for stage 3B lung cancer since August 2018.

Jump to this post

Hi @justapril, I moved your message to this existing discussion about COPD and stem cell therapy in the Lung Health group. I recommend reading through the past messages in this discussion as well as these:

Stem Cells / Regeneration https://connect.mayoclinic.org/discussion/stem-cells-regeneration/

Regenerative medicine for COPD is still in the research phase. Do use caution about promises made by some clinics.

To help people learn more about the proven therapies and the promise of developing therapies, Mayo Clinic offers a free telephone consult service. When you call the consult service, they will tell you about the availability of approved stem cell therapy at Mayo Clinic and elsewhere, and for what conditions. You can learn more about the Consult Service here http://www.mayo.edu/research/centers-programs/center-regenerative-medicine/patient-care/clinical-services/regenerative-medicine-consult-service. Or call 1-844-276-2003 to speak with one of our experts.

@justapril you may also wish to join the Lung Cancer group here: https://connect.mayoclinic.org/group/lung-cancer/

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COPD and stem cell treatment | Mayo Clinic Connect

Bone Marrow & Stem Cell Transplant | Weill Cornell Medicine

By Stem Cell Medicine

Bone Marrow & Stem Cell Transplant

The Bone Marrow and Stem Cell Transplant Program at Weill Cornell Medicine was established with the mission of providing the best care and most innovative research in a compassionate and comfortable environment.

We take a multidisciplinary approach to care for patients with cancer and blood diseases who need stem cell transplants, providing world-class clinical care in collaboration with experts in leukemia, lymphoma, myeloma and other blood disorders. Based at NewYork-Presbyterian/Weill Cornell Medical Center, one of the top ten general hospitals in the nation, the expertise of our consulting team is unsurpassed.

Our patients and families cope with life-threatening illness; as such, sensitivity and compassion are a priority for our team. We view each patient as an individual, and our approach ensures that each treatment regimen is narrowly tailored to meet the unique, changing needs of our patients and their families before, during and after transplant.

As New Yorks premier healthcare institution, Weill Cornell Medicine is at the forefront of scientific research and clinical trials, enabling us to provide a full range of diagnostic and treatment protocols, including the latest breakthroughs in medicine.

Our Team

Our team of internationally-recognized bone marrow transplant and stem cell surgery specialists is known for advanced work and published research in:

Treating patients with aggressive leukemia and myelodysplastic syndromes

Bridge protocols for patients with refractory lymphoma and leukemia

Novel strategies to mobilize stem cells and improve transplantation for patients with multiple myeloma, leukemia and lymphoma

Transplants for solid tumors, severe auto-immune disorders, and AIDS

Treatment

We pride ourselves on exceptional outcomes and offer patients the most advanced diagnostic methods and treatment therapies to improve quality of life, including:

Umbilical cord blood transplant

Outpatient transplant

Autologous stem cell transplant; uses stem cells extracted from the bone marrow or peripheral blood of the patients own blood

Allogeneic stem cell transplant; uses stem cells extracted from the bone marrow or peripheral blood of a matching donor

Hematopoietic stem cell transplant; used to treat certain cancers of the blood/bone marrow, including leukemia and myeloma

Matched unrelated donor stem cell transplantation through the National Donor Matching Program

Non-ablative "mini" transplants

Haplo-Cord Transplant, allowing us to find donors for all patients, regardless of age or ethnic background

Bendamustine, a therapy that is well-tolerated and has excellent response rates in patients with myeloma

Novel forms of transplant, offering hope and success to older patients with leukemia

Clinical Trials

Clinical trials are important to improve outcomes and offer new treatment options. At Weill Cornell Medicine, we conduct more studies in blood cancers than any of our regional peers, allowing us to provide our patients with access to many multi-phase clinical trials. As active members of the international cancer research community, our oncologists also collaborate with other research centers to offer patients the most promising treatments available.

Second Opinions

In concert with your referring physician, we are always available to offer a second opinion in the form of a consultation with one of our specialists.

Why Choose Us?

Our collaborative approach means our patients receive supportive, comprehensive care and the most cutting-edge stem cell therapy and treatments. This enables patients to receive the best possible transplant outcomes. Additionally, we offer more allogeneic stem cell transplants for older adults than any other center in New York City and the entire tri-state area.

For more information or to schedule an appointment, call us at 212-746-2119 or 212-746-2646.

Located in New York City, Weill Cornell Medical College is ranked among the nations best by U.S. News & World Report year after year.

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Bone Marrow & Stem Cell Transplant | Weill Cornell Medicine

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