Stem Cell Clinic

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Adult Stem Cell Therapy Abroad, Bangkok Stem Cell Clinic

By Stem Cell Treatment

Stem Cell Pool Re-population

In order to self-repair, living organisms have stem cells in central and peripheral locations which can be attracted to sites of injured tissues by alarm signals. In this way, these cells proliferate, migrate, and accumulate in those damaged sites. If this situation of alarm perpetuates, stem cells could be permanently exhausted from their original locations leading to irreversible disease. Basically, it could be a matter of stem cell quantity and effective availability at a certain time point when active regeneration is needed. Adult stem cell therapy re-populates the stem cell pool, this helps in fighting disease and improving the immune system.

The expected consequences of this situation could be the lack of an appropriate number of stem cells for further tissue replacement and regeneration and eventually the development of disease and ageing. For example, we could think that any alteration of this stem cell homoeostasis by constant and repetitive trauma, physical hyperactivity, chronic inflammation and chronic disease could provoke a persistent disequilibrium inside all these reserve locations. This could promote an irreversible and premature stem cell exhaustion, being impossible then for the organism to self-repair and survive. As we age we have less circulating stem cells. Introduction of new stem cells to our bodies circulation can improve health and repopulate our stem cell pool.

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Adult Stem Cell Therapy Abroad, Bangkok Stem Cell Clinic

Adult Stem Cell Therapy in Utah Docere Clinics

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Stem cells, specifically mesenchymal stem cells (MSCs), have been called patient-specific drug stores for injured tissues because of their broad range of healing abilities. MSCs are directly responsible for healing damaged tissues after injury. Upon encountering damaged tissue, they release proteins that decrease inflammation, kill invading microbes, and trigger the growth of new connective tissues and blood vessels.In the case of severe damage and cell death, MSCs have the ability to turn into healthy versions of damaged or destroyed cells that they encounter.

When we take MSCs from your own bone marrow, from your own fat, or from both, concentrate and/or isolate them, and then inject them directly into your problem area, we trick your body into thinking that there has been a new injury without actually causing any tissue insult, and you get a second chance at healing. In the case of advanced osteoarthritis where the population of stem cells has been depleted, we are repopulating the area with stem cells, and thereby restoring the bodys natural ability to heal itself.

Docere Clinics is an affiliate member of the Cell Surgical Network(CSN).It is the belief of the CSN that the most ethical approach to stem cell therapy is under the umbrella of IRB approved research protocols. An IRB is an Institutional Review Board;a committee responsible for approving and overseeing research on humans. IRBs are approved under the auspices of the U.S. Department of Human Research Protection. As such, our patients understand the investigational nature of our activities, are provided appropriate informed consents, and are followed continuously on an online database to chart their progress or any issues of concern. The efforts of the CSN will provide safety data,demonstrate effectiveness of treatments, and help to improve treatment programs going forward.

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Adult Stem Cell Therapy in Utah Docere Clinics

Stem Cell Treatment Jupiter FL | Alternative …

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Umbilical cord cells include stem cells, growth factors and a range of other beneficial proteins and compounds. We use blood from the umbilical cord which has been purified to get rid of any harmful substances that might cause rejection of the treatment by your body. We inject the treated cord blood into the affected area, where the various active compounds found in cord cells go to work immediately to begin inflammation reduction and the promotion of healthy cell division and renewal. Some of the active compounds at work include VEGF (Vascular Endothelial Growth Factor), IL-LRA (Interleukin-1, a receptor antagonist, stem cell factors (SCF), FGF-2 (Fibroblast Growth Factor-2) and Transforming Growth Factor-beta (TGF-beta). Each of these compounds has a slightly different effect, but the net result is that the damaged cells in your joints are given the ingredients they need to kick-start healthy renewal and regeneration. The injection changes the chemistry inside the joint, creating a healthier environment that encourages positive, healing changes to take place. A better blood supply to the area, a reduction in damaging chronic inflammation and stimulation of healthy tissue growth are all typical consequences of the minimally invasive stem cell treatments we provide. By using umbilical cord cells in this way, its possible to transform joint therapy into a holistic healing process that prompts the body to enhance its own regenerative efforts. This results in a natural process of joint health improvement in the weeks or months following the injection.

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Stem Cell Treatment Jupiter FL | Alternative ...

Cell Science, Stem Cell Research & Regenerative …

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Overview

Cell scienceis an emerging field of therapeutics and as well as stem cell therapyalso. In this Stem cells are playing widal role to develop into a newregenerative medicineof modern era in the globe. Most researchers believe thatStem Cellsare play a major role in future generations and give the big global business market across the world. This event leads us to build a great opportunity to fulfill the global needs and human welfare.

1.Molecular and Cellular Physiology and of Structural Biology

The cell structure is an important target structure for drugs and bacterial pathogens. It is composed of different protein filaments that are continuously remodeledto construct a dynamic cellular scaffold. The cytoskeleton is a scaffold that gives cells their diverse and adaptable shapes and that organizes their internal structures.The cytoskeleton plays a fundamental role in all aspects of cell mechanics, such as cell adhesion and motility, cell division, intracellular transport, the establishment of cell polarity and the organization of cells in tissues and organs.Many drugs and bacterial toxins act by blocking or activating cytoskeletal regulatory proteins.We primarily investigate the regulation of the cytoskeleton in the context of cancer and bacterial infections.

2.Cell Biology of Vertebrates, Microbes and Parasites

Vertebrates are multicellular, heterotrophic eukaryotes with tissues that develop from embryonic layers. Biologists have identified 1.3 million living species of animals. Estimates of the total number of animal species run far higher, from 10 to 20 million to as many as 100 to 200 million.in general vertebrates are all motile, heterotropic, and multicellular .Animals are ingestive heterotrophs unlike plants, who store their food as starch,and animals store their food as glycogen. Vertebrates cells lack of cell walls that provide structural support for plants and fungi .The multicellular bodies of animals are held together by extracellular structural proteins especially collagen. Vertebrate cells are made up of cells organized into tissues .each tissue specialized to some specific functions .vertebrates have their unique types of intracellular junctions, including tight junctions, desmosomes. And gap junctions together. Microbes are member of the group of eukaryotic organisms that includes unicellular microorganisms such as yeasts and molds, as well as multicellular fungi that produce familiar fruiting forms known as mushrooms. These organisms are classified as a kingdom, Fungi, which is separate from the other eukaryotic life kingdoms of plants and animals.

3.Current Research in Cell and Molecular Biology

Cell biology is playing a vital role in current scientific research oriented studies. Current situation is all about cell biology leads to invention of regenerative medicine and receptor and antibody mediated medicine. Stem cells are using as a therapy for diseases include bone marrow transplantation ,cancer therapy and treating in Alzheimers disease and cardiac treatments etc. The cell science research is mainly target to to achieve the diagnostic and therapeutic uses for the people across the global. Currently it has emerged as a rapidly diversifying field with the potential to address the worldwide organ shortage issue and comprises of tissue regeneration and organ replacement. Regenerative medicine save public health bodies money by reducing the need for long-term care and reducing associated disorders, with potential benefits for the world economy as a whole.

4.Nanotechnology: Stem Cells & Cancer

Nanotechnology in medicine offers some exciting possibilities. Some techniques are only imagined, while others are at various stages of testing, or actually being used today. Nanotechnologies in medicine involve applications of nano particles currently under development, as well as longer range research that involves the use of manufactured nano-robots to make repairs at the cellular level. Nanotechnology in medicine currently being developed involves employing nano particlesto deliver drugs, heat, light or other substances to specific types of cells (such as cancer cells). Particles are engineered so that they are attracted to diseased cells, which allow direct treatment of those cells. This technique reduces damage to healthy cells in the body and allows for earlier detection of disease.

5.Molecular and Cellular Basis of Growth and Regeneration

Cell growth is used in the biological cell development and cell reproduction. Where a cell, known as the mother cell, grows and divides to produce two daughter cells. In the cell development cytoplasmic and organelle volume increase and genetic material also. Regeneration is the process of renewal, restoration, and growth that makes genomes, cells,organisms, and ecosystems resilent to natural fluctuations or events that cause disturbance or damage.

6.Stem Cells, Self-Assembly, Tissue Growth and Regeneration

Stem cells have the remarkable potential to develop into many different cell types in the body during early life and growth. In addition, in many tissues they serve as a sort of internal repair system, dividing essentially without limit to replenish other cells as long as the person or animal is still alive. When a stem cell divides, each new cell has the potential either to remain a stem cell or become another type of cell with a more specialized function, such as a muscle cell, a red blood cell, or a brain cell.

7.Germline Stem Cells

Germ cells are specialized cells which are involved in reproduction. The most well-known examples of this type of cell are gametes, the sperm and eggs which come together to create a zygote which can develop into a foetus. In addition to gametes, a number of other cells involved in reproduction are also germ cells, including gonocytes, the cells which regulate the production of eggs and sperm. All germ cells carry the germ line, the genetic material which an organism can pass on to its offspring. In humans, these cells are haploid, meaning that they carry only half the number of chromosomes necessary to create an organism. When germ cells from two different people meet, their haploid genetic material combines to create diploid cells which can replicate themselves through cell division, ultimately building a baby.

8.Cellular And Molecular Medicine

Regenerative medicines have the ability to repair, replace, and regenerate tissues and organs affected due to injury, disease, or natural aging process. These medicines are capable of restoring the functionality of cells & tissues and are applicable in a wide range of degenerative disorders such as dermatology, Neurodegenerativediseases, cardiovascular and orthopaedic applications. Researchers focus on developing technologies based on biologics, genes, somatic as well as stem cells. Stem cells are capable of proliferation and differentiation owing to which they are of importance in this field.

9.Computational Biology and Drug Designing

Computational Biology is the science of using biological data to develop algorithms and relations among various biological systems. Computational biology is different from biological computation, which is a sub field of computer science and computer engineering using bio engineering and biology to build computers, but is similar to bioinformatics, which is an interdisciplinary science using computers to store and process biological data. Computer-aided drug design methods have played a major role in the development of therapeutically important small molecules for over three decades. The field is broadly defined and includes foundations in computer science, applied mathematics, animation, statistics, biochemistry, chemistry, biophysics, molecular biology, genetics, genomics, ecology, evolution, anatomy, neuroscience, and visualization.

10.Cell Signaling Technology

Cell signalling is major part of communication that coordinates basic activities of cells and perform cell actions. The ability of cells to perceive and correctly respond to their micro environment on the basis of development, tissue reform, and immunity as well as normal tissue homeostasis. Damage in cellular information processing are responsible for diseases such as cancer, autoimmunity, and diabetes. By understanding cell signalling, diseases may be treated more effectively and, theoretically, artificial tissues may be created. Cell signalling has been most studied in human diseases. Cell signalling may also occur between the cells of two different organisms. In mammals, early embryo cells exchange signals with cells of the uterus.

11.Tissue Engineering

Tissue engineering evolved from the field of bio material development and describes the practice of combining scaffolds, cells, and biologically active molecules into functional tissues. The goal of tissue engineering is to assemble functional constructs that restore, maintain, or improve damaged tissues or whole organs. Artificial skin and cartilage are examples of engineered tissues that have been approved by the FDA. This field continues to evolve. In addition to medical applications, non-therapeutic applications include using tissues as biosensorsto detect biological or chemical threat agents, and tissue chips that can be used to test the toxicity of an experimental medication.

12.Cell Rejuvenation and Wound Healing

Cell Rejuvenation is described as the reforming of a damaged cell. Skin compartments, epidermis, and hair follicles house stem cells that are indispensable for skin homeostasis and regeneration. The stem cells also contribute to wound repair, resulting in restoration of tissue integrity and function of damaged tissue. Unsuccessful wound healing processes often lead to non-healing wounds. Chronic wounds are caused by depletion of stem cells and a variety of other cellular and molecular mechanisms, many of which are still poorly understood. Current chronic wound therapies are limited, so the search to develop better therapeutic strategies is on going.Adult stem cells are gaining recognition as potential candidates for numerous skin pathologies. Emerging concepts offer some perspectives on how skin tissue-engineered products can be optimized to provide efficacious therapy in cutaneous repair and regeneration.

13.Stem Cell Therapeutics in Modern era

Stem-cell therapy is the use of stem cells to treat or prevent a disease or condition. Bone is the most widely used stem-cell therapy, but some therapies derived from umbilical cord blood are also in use. Research is underway to develop various sources for stem cells, and to apply stem-cell treatments for neurodegenerative diseases and conditions such as diabetes, heart disease, and other conditions. Stem-cell therapy has become controversial following developments such as the ability of scientists to isolate and culture embryonic stem cells, to create stem cells using somatic cell nuclear transfer and their use of techniques to create induced spluripotent stem cell. This controversy is often related to abortion politics and to human cloning. Additionally, efforts to market treatments based on transplant of stored umbilical cord blood have been controversial.

14.Cancer Cell Biology

Cancer stem cells are cancer cells that possess characteristics associated with normal stem cells, specifically the ability to give rise to all cell types found in a particular cancer sample. CSCs are therefore tumorogenic , perhaps in contrast to other non-tumorigenic cancer cells. CSCs may generate tumours through the stem cell processes of self-renewal and differentiation into multiple cell types. Such cells are hypothesized to persist in tumours as a distinct population and cause relapse and metastasis by giving rise to new tumours. Therefore, development of specific therapies targeted at CSCs holds hope for improvement of survival and quality of life of cancer patients, especially for patients with metastatic disease.

15.Bioethical Issues in Cell and Stem Cell Biology

The main Bioethical issues associated with human stem cells involve their derivation and use for research. Although there are interesting ethical issues surrounding the collection and use of somatic adult stem cells from aborted foetuses and umbilical cord blood, the most intense controversy to date has focused on the source of human embryonic stem (hES) cells. At present, new ethical issues are beginning to emerge around the derivation and use of other hES celllike stem cells that have the capacity to differentiate into all types of human tissue. In the near future, as the stem cell field progresses closer to the clinic, additional ethical issues are likely to arise concerning the clinical translation of basic stem cell knowledge into reasonably safe, effective and accessible patient therapies. This Review summarizes these and other bio ethical issues of the past, present and future of stem cell research.

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Cell Science, Stem Cell Research & Regenerative ...

Stem Cell Treatment for Optic Neuropathy – Beike Biotechnology

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Acupuncture

Acupuncture is a technique in which practitioners stimulate specific points on the body - most often by inserting thin needles through the skin. It is one of the most effective practices used in traditional Chinese medicine. Acupuncture stimulates nerve fibers to transmit signals to the spinal cord and brain, activating the bodys central nervous system. The spinal cord and brain then release hormones responsible for making us feel less pain while improving overall health. Acupuncture may also: increase blood circulation and body temperature, affect white blood cell activity (responsible for our immune function), reduce cholesterol and triglyceride levels, and regulate blood sugar levels.

Aquatherapy

Aquatic Physical Therapy is the practice of physical therapy in a specifically designed water pool with a therapist. The unique properties of the aquatic environment enhance interventions for patients with neurological or musculoskeletal conditions. Aquatic therapy includes a wide range of techniques allowing patients to improve their balance, muscle strength and body mechanics. Aquatic therapy works to enhance the rehabilitation process and support effectiveness of stem cell treatment.

Epidural Stimulation

Hyperbaric Oxygen Therapy

Hyperbaric Oxygen Therapy (HBOT) is the medical use of oxygen at a level higher than atmospheric pressure. The equipment required consists of pressure chamber, which may be of rigid or flexible construction, and a means of delivering 100% oxygen into the respiratory system. Published research shows that HBOT increases the lifespan of stem cells after injection and provides an oxygen-rich atmosphere for the body to function at optimum levels.

Nerve Growth Factor (NGF)

Nerve growth factor (NGF) is a member of the neurotrophic factor (neurotrophin, NTFS) family, which can prevent the death of nerve cells and has many features of typical neurotransmitter molecules. NGF plays an important role in the development and growth of nerve cells. NGF is synthesized and secreted by tissues (corneal epithelial, endothelial, and corneal stromal cells), and it can be up-taken by sympathetic or sensory nerve endings and then transported to be stored in neuronal cell bodies where it can promote the growth and differentiation of nerve cells.NGF can exert neurotrophic effects on injured nerves and promote neurogenesis (the process of generating neurons from stem cells) that is closely related to the development and functional maintenance and repair of the central nervous system. It is also capable of promoting the regeneration of injured neurons in the peripheral nervous system, improving the pathology of neurons and protecting the nerves against hypoxia (lack of oxygen)/ischemia (lack of blood supply).

Nutrition Therapy

Occupational Therapy

Occupational therapy interventions focus on adapting the environment, modifying the task and teaching the skill, in order to increase participation in and performance of daily activities, particularly those that are meaningful to the patient with physical, mental, or cognitive disorders. Our Occupational Therapists also focus much of their work on identifying and eliminating environmental barriers to independence and participation in daily activities, similar to everyday life.

Physiotherapy

Physical therapy or physiotherapy (often abbreviated to PT) is a physical medicine and rehabilitation specialty that, by using mechanical force and movements, remediates impairments and promotes mobility, function, and quality of life through examination, diagnosis, prognosis, and physical intervention. We combine our PT with stem cells for maximum physical rehabilitation improvements.

Transcranial Magnetic Stimulation

Research has shown that TMS can effectively treat symptoms of depression, anxiety, neurological pain, stroke, spinal cord injuries, autism and more. This procedure is very simple and noninvasive. During the procedure, a magnetic field generator or coil is placed near the head of the person receiving the treatment. The coil produces small electrical currents in the region of the brain just under the coil via electromagnetic induction. This electrical field causes a change in the transmembrane current of the neuron which leads to depolarization or hyperpolarization of the neuron and the firing of an action potential.

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Stem Cell Treatment for Optic Neuropathy - Beike Biotechnology

Cell Therapy – Celgene

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Celgene Cellular Therapeutics (CCT) is at the forefront of cell therapy research and development. Our lead cell therapy candidates are derived from healthy, full-term placenta, a remarkable organ shared by mother and baby. From this immunologically privileged source, we have generated novel cell therapy candidates with immunomodulatory, anti-inflammatory, angiogenic and reparative properties.

CCTs placenta-derived adherent cells (PDAC cells) have been extensively characterized (see our Publications) and display multiple activities including the ability to stimulate natural repair processes, with the potential to transform the treatment of a broad range of serious debilitating diseases including autoimmune, vascular and neurodegenerative disorders. Specific formulations of PDAC cells are our lead clinical candidates PDA-001 (intravenous) and PDA-002 (intramuscular), currently in clinical trials for Crohns disease, Peripheral Artery Disease with Diabetic Foot Ulcer and other indications.

Cell therapy research teams focus on the discovery and early development of novel cell-based therapeutics. We have developed a portfolio of unique clinical candidates with broad therapeutic potential, including human placenta-derived stem cells (HPDSC), natural killer (NK) cells and amnion-derived adherent cells (AMDAC cells). We are industry leaders in adult stem cell isolation, cell culture, characterization, functional interpretation and translational medicine. Our state-of-the-art in vitro, preclinical and translational biology approaches conducted in-house and in collaboration with international expert groups, are specifically designed to elucidate the complex mechanisms that are associated with live cell-based therapeutics

We are constantly searching for new ways to bring innovative solutions to unmet medical needs. Celgene is a leader in cancer treatments, and we are investigating novel cell therapies to expand this arsenal, both in-house and through alliances with other leaders. Our collaborations with bluebird bio and Baylor College of Medicine on chimeric antigen receptor-modified T-cell-based (CAR T) therapies genetically modifying a patients own cells to fight cancer have the potential to revolutionize patient care in a range of hematological and solid malignancies.

CCT is proud to work with federal research agencies and has been a performer for the Defense Advanced Research Project Agency (DARPA) since 2008. CCT has led two international consortia to develop advanced stem cell manufacturing and differentiation methods and hematopoietic stem cell-based therapeutic concepts and product candidates, leading to breakthrough findings in these areas.

CCT is a world leader in cell therapy process development and clinical manufacturing. We possess in-house GMP facilities and development laboratories for processing allogeneic and autologous cells from a variety of cell sources while maintaining the highest level of regulatory compliance. We have broad experience in developing, optimizing, scaling up and validating cell therapy processes and assays, from donor tissue procurement through product supply and distribution. Our integrated Technical Operations laboratories and staff share the same location, enabling us to gain process understanding from production operations and to translate development activities into manufacturing science. In summary, CCT Technical Operations has all systems, personnel, facilities, and capabilities necessary for taking a cell therapy product from discovery into the clinic and to commercialization.

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Cell Therapy - Celgene

Meet Dr. Garg, M.D., Experienced Stem Cell Doctor

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Meet the Leading Expert in Regenerative Medicine (Autologous Adult Stem Cell Treatments) For Non-Surgical ALTERNATIVES to Your Orthopedic Conditions, Such as Problems of the Spine, Hip, Knee, Shoulder, & Other Joints from Arthritis, Sports Injuries and other common medical conditions.

Hardesh Kumar Garg, M.D., is pioneering effective and safe, non-surgical alternatives to orthopedic and sports surgery.

He is a leading physician in the United States specializing in emerging and exciting field of Adult Stem Cell and PRP treatments, Anti-Aging Medicine and Sports Medicine.

Dr. Garg is one of the few physicians in the United States who are trained in this specialized field of Regenerative Medicine. He uses your own (Autologous) stem cells from Bone Marrow, and Fat; along with Platelet-Rich Plasma (PRP) and anti-aging hormone replacement protocols. He has also designed specialized supplements to boost immune system and decrease chronic bad inflammation in your body, thus increasing the success of his stem cell/regenerative medicine procedures for his patients.

He has helped many patients from around the country by treating their pain and injuries with his unique clinical protocols combining Bone Marrow Stem Cells, Adipose (Fat) Derived Stem Cells and Platelet-Rich Plasma (PRP), while boosting their immune and hormone systems with hormone balancing protocols and nutritional/herbal supplementations.

People suffering from back and neck pain, sports and auto injuries, arthritis or other serious pain often fear visiting doctors because they dont want to hear the word surgery. The fear is even greater among senior citizens as the healing process may take longer, complications can be more dramatic and results more difficult to perceive.

Dr. Garg provides high quality, cutting-edge, proven, safe and cost-effective alternatives to dangerous and costly orthopedic joint, tendon and spine surgeries.

He has served as clinical assistant professor at the Department of Internal Medicine at the University of Florida School of Medicine and has also worked as a courtesy clinical assistant professor at the University of South Florida School of Medicine.

Dr. Garg has experience working with professional and amateur athletes from different sports to increase their performance with this unique and individualized health and wellness program. He has worked with professional athelets like former number one PGA Tour Golfer, several NFL players, many Tennis athletes and athletes from various other sports.

Affectionately known as Dr. G to his patients, an internal medicine specialist, received his medical degree from the University of Delhi, India and completed post-graduate training in internal medicine from West Virginia University Health Sciences Center (Morgantown, WV) in 1993.

As a clinician, researcher, teacher, scientist and writer, Dr. G has been practicing medicine for more than 20 years.

He worked as sub-investigator in a National Institute of Health-funded research study seeking genetic information on obesity.

He has served as a member of the Medical Advisory Board of H Magazine, a publication of The Florida Times-Union, Jacksonville's daily newspaper.

He also served as the national medical advisor for a national chain of weight loss centers.

He was a speaker for major International Pharmaceutical companies like Pfizer and Bristol- Myers-Squibb.

Dr. G is a member of Duval County Medical Society, American Academy of Pain Medicine, American Academy of Anti-Aging Medicine and American Society for Laser Medicine and Surgery.

Publications:

Association analyses of adrenergic receptor polymorphisms with obesity and metabolic alterations - John J. Lima(a), Hua Feng(b), Laurie Duckworth(a), Jianwei Wang(a), James E. Sylvester(a), Niranjan Kissoon(c),Hardesh Garg(d).

Metabolism Clinical and Experimental 56 (2007) 757 765, Received 11 September 2006; accepted 2 January 2007

Professional Presentations:

Clinical Investigator:

Principal Clinical Investigator, Clinical Research:

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Meet Dr. Garg, M.D., Experienced Stem Cell Doctor

Induced Pluripotent Stem Cells | The Progeria Research Foundation

By Induced Pluripotent Stem Cells

The Progeria Research Foundation Cell & Tissue BankHuman Induced Pluripotent Stem Cells (iPSC)

1. iPSC Background information for the non-scientist Stem cells are immature cells that have not yet committed to becoming any one cell type. They are pliable because they have the potential to develop into many different types of mature cells in the body, such as cells that make up the heart or blood vessels, and other tissues and organs. In 2007, researchers discovered a strategy for creating stem cells in the laboratory by reprogramming mature adult cells that we commonly grow for research purposes.1, 2 . These artificially created stem cells are called Induced Pluripotent Stem Cells (iPSCs). For the field of Progeria, this is a huge breakthrough. For the first time, scientists can now make Progeria stem cells and ask questions about how stem cells function and develop in Progeria. Previously there was no source of human Progeria stem cells, and there was therefore a void of information about how Progeria stem cells function compared with stem cells from people without Progeria. In addition, scientists can re-program the Progeria stem cells to create, for the first time, mature Progeria blood vessels, heart cells, and other cell types. Until now, there was no source of human Progeria heart or blood vessel cells. We can now ask key questions about the heart disease that leads to early death in Progeria from heart attacks and strokes. We can compare these discoveries with the heart disease and aging in the general population and discover more about what influences aging in all of us. Already there have been several excellent studies published using Progeria stem cells.3-5 Our goal at The Progeria Research Foundation is to facilitate many more discoveries using this invaluable tool. For a primer on stem cells, please see this US government website: http://stemcells.nih.gov/info/basics/defaultpage.asp

2.Purpose of induced pluripotent stem cell (iPSC) generation and distribution by The Progeria Research Foundation The mission of The Progeria Research Foundation is to discover treatments and the cure for Hutchinson-Gilford Progeria Syndrome and its aging-related disorders. In 2009, PRF entered into a collaboration with an expert team of scientists at the University of Toronto, Canada, under the direction of William Stanford, PhD, to generate high quality Progeria iPSCs. Dr. Stanford is the Canada Research Chair in Integrative Stem Cell Biology. As of 2011, PRF continues to collaborate with Dr. Stanford at the University of Ottawa, Canada where he is Professor of Cellular and Molecular Medicine, Faculty of Medicine, and Senior Scientist at Ottawa Hospital Research Institutes Sprott Centre for Stem Cell Research.

Our goal is to provide this invaluable tool to researchers throughout the world. This new research tool will be used to generate new and innovative research in Progeria, as well as its relationship to heart disease and aging.

3. Generation of Hutchinson-Gilford Progeria Syndrome Induced-Pluripotent Stem Cells (iPSCs) Induced-Pluripotent Stem Cells (iPSCs) were derived using VSVG-pseudotyped retroviral transduction of four human factors, Oct4, Sox2, Klf4, and c-Myc into fibroblasts. iPSC colonies were derived on mouse-embryonic fibroblasts (MEFs). The procedure used was essentially as previously described but without the use of the EOS reporter (Nature Protocols 4: 1828-1844, 2009).

4. Quality Control: Validation and Characterization The lines that are currently available have undergone several validation steps (see downloadable PDFs below):

Additional validation in process: Some lines have completed teratoma assays as shown in supporting data. For all other lines, teratoma assays are in process and status will be updated as these assays are completed.

5. Original starting material from which these iPS cells were derived iPSCs were derived from PRF Cell & Tissue Bank non-transformed fibroblast cell lines.

The transduction method used for all iPS lines was Retrovirus MKOS.

iPSC Line ID

Mutation

Gender and Donation Age

HGADFN003 iPS 1B

LMNAExon 11, 1824 C>T

Male 2yr 0mo

Dermal Fibroblasts HGADFN003

HGADFN003 iPS 1C

LMNA Exon 11, 1824 C>T

Male 2yr 0mo

Dermal Fibroblasts HGADFN003

HGDFN003 iPS 1D

LMNA Exon 11, 1824 C>T

Male 2yr 0mo

Dermal Fibroblasts HGADFN003

HGADFN167 iPS 1J

LMNA Exon 11, 1824 C>T

Male 8yr 5mo

Dermal Fibroblasts HGADFN167

HGADFN167 iPS 1Q

LMNA Exon 11, 1824 C>T

Male 8yr 5mo

Dermal Fibroblasts HGADFN167

HGMDFN090 iPS 1B

Mother of HGADFN167 (unaffected)

Female 37yr 10mo

Dermal Fibroblasts HGMDFN090

HGMDFN090 iPS 1C

Mother of HGADFN167 (unaffected)

Female 37yr 10mo

Dermal Fibroblasts HGMDFN090

HGFDFN168 iPS1 D2

Father of HGADFN167 (unaffected)

Male 40yr 5mo

Dermal Fibroblasts HGFDFN168

HGFDFN168 iPS1P

Father of HGADFN167 (unaffected)

Male 40yr

5mo

Dermal Fibroblasts HGFDFN168

PRF AVAILABLE CELL LINES

6. Join our email list for future iPSC updates and new cell lines We are continuing to generate iPSC lines. If you would like periodic updates on iPSCs held in the PRF Cell & Tissue Bank,please join our emailing list by clicking here

7. Questions? Please contact Leslie Gordon, MD, PhD, Medical Director, with any questions or needs, at lgordon@progeriaresearch.org or 978-535-2594

8. Ordering iPS cell lines

In 2014, PRF instituted a policy of no changes to our MTA. This is the result of 12 years of contractual arrangements with 70 research teams working at institutions in 14 countries. PRF and its counsel have taken into consideration the issues that have arisen in that time period and edited the agreement accordingly, resulting in what we feel are fair and reasonable terms.

For U.S. Federal Government Institutions, please contact Joan Brazier, Research Study Coordinator, at joan_brazier@brown.edu or 401-863-9628.

Step 1: Complete an application and material transfer agreement Application and Agreement for Non-government Institutions

Material Transfer Agreement for Non-government Institutions*

Step 2: Return the completed application and material transfer agreement to PRF at info@progeriaresearch.org. Once approved, you will receive an email confirming your order and anticipated shipping date.

Step 3: Dr. Stanfords laboratory is currently distributing lines as live cultures. His laboratory will email you when the culture has been shipped, with shipping and tracking information. Inexperienced researchers are directed to obtain training at specialized courses essential to human embryonic stem cell/iPSCs work.

Step 4: The University of Ottawa will charge $84.00 per iPSC line plus courier costs, if any, and will send you a bill directly.

9. HGPS and Control iPS Cell Culture Media Preparation Culturing Progeria iPSCs requires the preparation of various kinds of media depending on the growth conditions of the cells and the experimental requirements. In addition to maintenance media, there is also supportive media for the MEFs. The HGPS iPSCs were derived using a Knock-Out medium containing Knock Out Serum Replacement (KOSR).

MEF medium

Store at 4C and use within 4 weeks. If purchasing untreated MEFs from Millipore it is recommended to increase the FBS concentration to 20% for better growth during expansion.

HGPS and Control hiPSC media

Reagent

We recommend Lot testing the Knockout Serum Replacement on established hES cells before being used for Progeria iPS cells.

10. Preparation of HGPS and Control iPSC Culture Surfaces To maintain high quality cells and colonies, it is imperative to passage onto appropriate surfaces. This surface could consist of inactivated mouse embryonic fibroblasts (MEFs, replication arrested through irradiation or mitomycin-c treatment). The protocol for inactivation of MEFs by irradiation follows. However MEFs can also be inactivated by treatment with mitomycin C if there is no access to an irradiator. Inactivated MEFs can be made in house or purchased through Millipore (cat# PMEF-CFL for MEFs that have not been mitotically inactivated or cat# PMEF-CFL for inactivated ones that are ready to use). A vial of untreated MEFs can be expanded and treated with Mitomycin C used immediately or frozen down for future use.

11. Inactivating (by irradiation) and plating MEFs

Materials:

Procedure:

12. Thawing HGPS and Control iPS cell lines One vial of hiPSCs should be thawed into one well of a 6-well plate containing inactivated mouse embryonic feeders cells (MEFs).

Have all tubes, warmed medium, and plates ready before starting the protocol to ensure that the thawing procedure is done as quickly as possible.

Materials:

Procedure:

Note: If only a few undifferentiated colonies are observed after thawing, it may be necessary to select only these colonies for passaging and replate them in the same size well on a new plate.

13. Routine Passaging and Maintenance of Undifferentiated HGPS and Control iPSCs

In order to assure healthy cells, it is important to change their media on a daily basis. This is a simple process of aspirating the old media and replacing it with fresh iPS media. After some time, usually 4-6 days after splitting, it will be necessary to split the cells once again. Splitting cells before they become too confluent will ensure a higher number of undifferentiated cells. Usually a 1:6 or 1:8 split will work well and allow 6-7 days between passages.

14. Suggested Protocol for Passaging iPS CellsUpdated September 4, 2014

The following protocol, obtained from Beers et al, 2012 has been giving excellent results for the team at the Human Pluripotent Stem Cell Facility of Ottawa Hospital Research Institute. According to this team, the protocol has dramatically helped to decrease the differentiated cells that might start to grow and it speeds up the passaging. Cells are often ready within 3 to 5 days instead of 5 to 7. Therefore this could save time and money on media.

EDTA solution: Add 500ul of 0.5M EDTA (pH 8.0) into 500ml of DPBS (-/-). Add 0.9g of NaCl and adjust the osmolarity to 340 mOsm. Filter the solution to sterilize and store it at 4C for up to 6 months. The goal is to create the least amount of disturbance for the cells during dissociation. Therefore the EDTA solution is at the same osmolarity as the E8 media.

Protocol

Add 2ml of E8 media to a 6 well matrigel coated plate.

Take the plate to be passaged and remove the media from the well and wash twice with 1ml of PBS(-/-).

Add 1ml of the EDTA solution to the well and leave for 4min at room temperature.

Once 4 min. is up remove EDTA solution and add 1ml of E8 media.

Scrape cells and divide cells amongst the 6 wells of your plate containing E8 media (Ive been taking 160ul into each well). Avoid breaking up the pieces as much as possible. Preferably use a wide mouth pipette tip.

Swirl and incubate at 37C.

NOTE: Once the cells have been scraped, transfer them to the new plate as soon as possible because the cells will re-attach quickly.

Source:

Passaging and colony expansion of human pluripotent stem cells by enzyme-free dissociation in chemically defined culture conditions

Jeanette Beers, Daniel R. Gulbranson, Nicole George, Lauren I. Siniscalchi, Jeffrey Jones, James A. Thomson, and Guokai Chen

Nat Protoc. 2012 Nov; 7(11):2029-40

15. Culturing HGPS and Control iPSCs on MEFs

Materials:

Procedure:

16. Cryopreservation of HGPS and Control iPSCs

Multiple passaging and expansion of iPSCs will result in a surplus of cells. Instead of disposing, it is good practice to freeze cells on occasion to build up a stock and give you cells you can go back to and thaw out for use in the future.

The protocols described below are based on iPSC cultures in 6-well plates where initial clump seeding is adjusted so that wells are 60 70% confluent at time of cryopreservation. Before cryopreservation, iPSCs should be of high quality (primarily undifferentiated with less than 20% of the cells being differentiated). Cryopreservation should be done approximately 1 day before the cells are ready to passage. iPSCs will have improved survival following thawing if cryopreserved as large clumps.

Link:
Induced Pluripotent Stem Cells | The Progeria Research Foundation

Induced Pluripotent Stem Cells for Cardiovascular Diagnostics

By Induced Pluripotent Stem Cells

Nearly 500,000 people in the US die of sudden cardiac death each year, and long QT syndrome (LQTS) is a major form of sudden cardiac death. LQTS can be triggered by drug exposure or stresses. Drug-induced LQTS is the single most common reason for drugs to be withdrawn from clinical trials, causing major setbacks to drug discovery efforts and exposing people to dangerous drugs. In most cases, the mechanism of drug-induced LQTS is unknown. However, there are genetic forms of LQTS that should allow us to make iPS cellderived heart cells that have the key features of LQTS. Our objective is to produce a cell-based test for LQTS with induced pluripotent stem (iPS) cell technology, which allows adult cells to be reprogrammed to be stem celllike cells. Despite the critical need, current tests for drug-induced LQTS are far from perfect. As a result, potentially unsafe drugs enter clinical trials, endangering people and wasting millions of dollars in research funds. When drugs that cause LQTS, such as terfenadine (Seldane), enter the market, millions of people are put at serious risk. Unfortunately, it is very difficult to know when a drug will cause LQTS, since most people who develop LQTS have no known genetic risk factors. The standard tests for LQTS use animal models or hamster cells that express human heart genes at high levels. Unfortunately, cardiac physiology in animal models (rabbits and dogs) differs from that in humans, and hamster cells lack many key features of human heart cells. Human embryonic stem cells (hESCs) can be differentiated into heart cells, but we do not know the culture conditions that would make the assay most similar to LQTS in a living person. These problems could be solved if we had a method to grow human heart cells from people with genetic LQTS mutations, so that we know the exact test conditions that would reflect the human disease. This test would be much more accurate than currently available tests and would help enable the development of safer human pharmaceuticals. Our long-term goal is to develop a panel of iPS cell lines that better represent the genetic diversity of the human population. Susceptibility to LQTS varies, and most people who have life-threatening LQTS have no known genetic risk factors. We will characterize iPS cells with well-defined mutations that have clinically proven responses to drugs that cause LQTS. These iPS cell lines will be used to refine testing conditions. To validate the iPS cellbased test, the results will be directly compared to the responses in people. These studies will provide the foundation for an expanded panel of iPS cell lines from people with other genetic mutations and from people who have no genetically defined risk factor but still have potentially fatal drug-induced LQTS. This growing panel of iPS cell lines should allow for testing drugs for LQTS more effectively and accurately than any current test. To meet these goals, we made a series of iPS cells that harbor different LQTS mutations. These iPS cells differentiate into beating cardiomyocytes. We are now evaluating these LQTS cell lines in cellular assays. We are hopeful that our studies will meet or exceed all the aims of our original proposal.

Read the original here:
Induced Pluripotent Stem Cells for Cardiovascular Diagnostics

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