Stem Cell Clinic

Keeping stem cells pluripotent

By Stem Cell Medicine

PUBLIC RELEASE DATE:

13-Jan-2014

Contact: Scott LaFee slafee@ucsd.edu 619-543-6163 University of California - San Diego

While the ability of human embryonic stem cells (hESCs) to become any type of mature cell, from neuron to heart to skin and bone, is indisputably crucial to human development, no less important is the mechanism needed to maintain hESCs in their pluripotent state until such change is required.

In a paper published in this week's Online Early Edition of PNAS, researchers from the University of California, San Diego School of Medicine identify a key gene receptor and signaling pathway essential to doing just that maintaining hESCs in an undifferentiated state.

The finding sheds new light upon the fundamental biology of hESCs with their huge potential as a diverse therapeutic tool but also suggests a new target for attacking cancer stem cells, which likely rely upon the same receptor and pathway to help spur their rampant, unwanted growth.

The research, led by principal investigator Karl Willert, PhD, assistant professor in the Department of Cellular and Molecular Medicine, focuses upon the role of the highly conserved WNT signaling pathway, a large family of genes long recognized as a critical regulator of stem cell self-renewal, and a particular encoded receptor known as frizzled family receptor 7 or FZD7.

"WNT signaling through FZD7 is necessary to maintain hESCs in an undifferentiated state," said Willert. "If we block FZD7 function, thus interfering with the WNT pathway, hESCs exit their undifferentiated and pluripotent state."

The researchers proved this by using an antibody-like protein that binds to FZD7, hindering its function. "Once FZD7 function is blocked with this FZD7-specific compound, hESCs are no longer able to receive the WNT signal essential to maintaining their undifferentiated state."

FZD7 is a so-called "onco-fetal protein," expressed only during embryonic development and by certain human tumors. Other studies have suggested that FZD7 may be a marker for cancer stem cells and play an important role in promoting tumor growth. If so, said Willert, disrupting FZD7 function in cancer cells is likely to interfere with their development and growth just as it does in hESCs.

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Keeping stem cells pluripotent

Nature Study Discovers Chromosome Therapy to Correct a Severe Chromosome Defect

By Stem Cell Medical Center

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Newswise Geneticists from Ohio, California and Japan joined forces in a quest to correct a faulty chromosome through cellular reprogramming. Their study, published online today in Nature, used stem cells to correct a defective ring chromosome with a normal chromosome. Such therapy has the promise to correct chromosome abnormalities that give rise to birth defects, mental disabilities and growth limitations.

In the future, it may be possible to use this approach to take cells from a patient that has a defective chromosome with multiple missing or duplicated genes and rescue those cells by removing the defective chromosome and replacing it with a normal chromosome, said senior author Anthony Wynshaw-Boris, MD, PhD, James H. Jewell MD '34 Professor of Genetics and chair of Case Western Reserve School of Medicine Department of Genetics and Genome Sciences and University Hospitals Case Medical Center.

Wynshaw-Boris led this research while a professor in pediatrics, the Institute for Human Genetics and the Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research at UC, San Francisco (UCSF) before joining the faculty at Case Western Reserve in June 2013.

Individuals with ring chromosomes may display a variety of birth defects, but nearly all persons with ring chromosomes at least display short stature due to problems with cell division. A normal chromosome is linear, with its ends protected, but with ring chromosomes, the two ends of the chromosome fuse together, forming a circle. This fusion can be associated with large terminal deletions, a process where portions of the chromosome or DNA sequences are missing. These deletions can result in disabling genetic disorders if the genes in the deletion are necessary for normal cellular functions.

The prospect for effective counter measures has evaded scientistsuntil now. The international research team discovered the potential for substituting the malfunctioning ring chromosome with an appropriately functioning one during reprogramming of patient cells into induced pluripotent stem cells (iPSCs). iPSC reprogramming is a technique that was developed by Shinya Yamanaka, MD, PhD, a co-corresponding author on the Nature paper. Yamanaka is a senior investigator at the UCSF-affiliated Gladstone Institutes, a professor of anatomy at UCSF, and the director of the Center for iPS Cell Research and Application (CiRA) at the Institute for Integrated Cell-Material Sciences (iCeMS) in Kyoto University. He won the Nobel Prize in Medicine in 2012 for developing the reprogramming technique.

Marina Bershteyn, PhD, a postdoctoral fellow in the Wynshaw-Boris lab at UCSF, along with Yohei Hayashi, PhD, a postdoctoral fellow in the Yamanaka lab at the Gladstone Institutes, reprogrammed skin cells from three patients with abnormal brain development due to a rare disorder called Miller Dieker Syndrome, which results from large terminal deletions in one arm of chromosome 17. One patient had a ring chromosome 17 with the deletion and the other two patients had large terminal deletions in one of their chromosome 17, but not a ring. Additionally, each of these patients had one normal chromosome 17.

The researchers observed that, after reprogramming, the ring chromosome 17 that had the deletion vanished entirely and was replaced by a duplicated copy of the normal chromosome 17. However, the terminal deletions in the other two patients remained after reprogramming. To make sure this phenomenon was not unique to ring chromosome 17, they reprogrammed cells from two different patients that each had ring chromosomes 13. These reprogrammed cells also lost the ring chromosome, and contained a duplicated copy of the normal chromosome 13.

It appears that ring chromosomes are lost during rapid and continuous cell divisions during reprogramming, said Yamanaka. The duplication of the normal chromosome then corrects for that lost chromosome.

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Nature Study Discovers Chromosome Therapy to Correct a Severe Chromosome Defect

Study discovers chromosome therapy to correct severe chromosome defect

By Stem Cell Medical Center

Jan. 13, 2014 Geneticists from Ohio, California and Japan joined forces in a quest to correct a faulty chromosome through cellular reprogramming. Their study, published online today in Nature, used stem cells to correct a defective "ring chromosome" with a normal chromosome. Such therapy has the promise to correct chromosome abnormalities that give rise to birth defects, mental disabilities and growth limitations.

"In the future, it may be possible to use this approach to take cells from a patient that has a defective chromosome with multiple missing or duplicated genes and rescue those cells by removing the defective chromosome and replacing it with a normal chromosome," said senior author Anthony Wynshaw-Boris, MD, PhD, James H. Jewell MD '34 Professor of Genetics and chair of Case Western Reserve School of Medicine Department of Genetics and Genome Sciences and University Hospitals Case Medical Center.

Wynshaw-Boris led this research while a professor in pediatrics, the Institute for Human Genetics and the Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research at UC, San Francisco (UCSF) before joining the faculty at Case Western Reserve in June 2013.

Individuals with ring chromosomes may display a variety of birth defects, but nearly all persons with ring chromosomes at least display short stature due to problems with cell division. A normal chromosome is linear, with its ends protected, but with ring chromosomes, the two ends of the chromosome fuse together, forming a circle. This fusion can be associated with large terminal deletions, a process where portions of the chromosome or DNA sequences are missing. These deletions can result in disabling genetic disorders if the genes in the deletion are necessary for normal cellular functions.

The prospect for effective counter measures has evaded scientists -- until now. The international research team discovered the potential for substituting the malfunctioning ring chromosome with an appropriately functioning one during reprogramming of patient cells into induced pluripotent stem cells (iPSCs). iPSC reprogramming is a technique that was developed by Shinya Yamanaka, MD, PhD, a co-corresponding author on the Nature paper. Yamanaka is a senior investigator at the UCSF-affiliated Gladstone Institutes, a professor of anatomy at UCSF, and the director of the Center for iPS Cell Research and Application (CiRA) at the Institute for Integrated Cell-Material Sciences (iCeMS) in Kyoto University. He won the Nobel Prize in Medicine in 2012 for developing the reprogramming technique.

Marina Bershteyn, PhD, a postdoctoral fellow in the Wynshaw-Boris lab at UCSF, along with Yohei Hayashi, PhD, a postdoctoral fellow in the Yamanaka lab at the Gladstone Institutes, reprogrammed skin cells from three patients with abnormal brain development due to a rare disorder called Miller Dieker Syndrome, which results from large terminal deletions in one arm of chromosome 17. One patient had a ring chromosome 17 with the deletion and the other two patients had large terminal deletions in one of their chromosome 17, but not a ring. Additionally, each of these patients had one normal chromosome 17.

The researchers observed that, after reprogramming, the ring chromosome 17 that had the deletion vanished entirely and was replaced by a duplicated copy of the normal chromosome 17. However, the terminal deletions in the other two patients remained after reprogramming. To make sure this phenomenon was not unique to ring chromosome 17, they reprogrammed cells from two different patients that each had ring chromosomes 13. These reprogrammed cells also lost the ring chromosome, and contained a duplicated copy of the normal chromosome 13.

"It appears that ring chromosomes are lost during rapid and continuous cell divisions during reprogramming," said Yamanaka. "The duplication of the normal chromosome then corrects for that lost chromosome."

"Ring loss and duplication of whole chromosomes occur with a certain frequency in stem cells," explained Bershteyn. "When chromosome duplication compensates for the loss of the corresponding ring chromosome with a deletion, this provides a possible avenue to correct large-scale problems in a chromosome that have no chance of being corrected by any other means."

"It is likely that our findings apply to other ring chromosomes, since the loss of the ring chromosome occurred in cells reprogrammed from three different patients," said Hayashi.

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Study discovers chromosome therapy to correct severe chromosome defect

DaSilva Institute Brings World-Class Medicine to Sarasota, Florida

By Stem Cell Medical Center

Sarasota, FL (PRWEB) January 13, 2014

The DaSilva Institute opened their brand new state-of-the-art medical facility in Sarasota, Florida on December 16, 2013.

The DaSilva Institute combines functional medicine with anti-aging and regenerative medicine, making it the most unique multi-specialty medical center of its kind in the U.S.

One major advantage that the DaSilva Institute has over similar centers found elsewhere in the US and overseas is its focus on autologous stem cell therapy. Used to reverse degenerative diseases and injuries, this innovative therapy involves harvesting stem cells from the patients own body fat without the controversial use of embryos, umbilical cords, placentas or donors, thus eliminating the risk of viruses and rejection.

The DaSilva Institute is also known for their expertise in bio-identical hormone replacement therapies, functional gastrointestinal disorders, mood disorders, nutritional counseling, IV nutrition and chelation, natural cancer support, regenerative orthopedics, platelet rich plasma (PRP), prolotherapy, and several new aesthetic treatments including facial rejuvenation, natural breast and buttocks augmentation and gentle liposculpture.

Guy DaSilva, MD, founder and medical director of the DaSilva Institute, states, Our vision is to make this extraordinary form of medicine accessible and affordable for people in the U.S. You shouldnt have to fly to other countries and spend tens of thousands of dollars for what you can receive in your own backyard for much less.

After outgrowing their previous office in the Lakewood Ranch area, the decision to move into a larger, more optimally equipped facility led them to the heart of Sarasota.

Dr. DaSilva states, My hope is that people will benefit from our extended menu of services and enjoy the beautiful and comforting ambiance of our new office, as well as the convenience of the new Sarasota location. And above all, we want to help more people discover health without limits.

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DaSilva Institute Brings World-Class Medicine to Sarasota, Florida

T-Cell Finding Sheds Light on Why HIV Can Persist Despite Treatment

By Stem Cell Treatment

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Newswise Ryan Zurakowski, assistant professor of electrical and computer engineering at the University of Delaware, is co-author of a paper appearing in Nature Medicine on Jan. 12 highlighting the role of T-cells in HIV.

The paper, titled HIV-1 Persistence in CD4+ T-Cells with Stem Cell-Like Properties, provides evidence that a particular T-cell type may help researchers better understand why HIV can persist despite treatment.

Zurakowskis co-authors include Mathias Lichterfeld, the papers lead author, and researchers from Massachusetts General Hospital (MGH); Ragon Institute of MGH, the Massachusetts Institute of Technology and Harvard University; the First Affiliated Hospital of China Medical University; Brigham and Womens Hospital; and Howard Hughes Medical Institute.

Zurakowski explained that HIV treatments do not kill infected cells. Instead, they stop the infection of new cells, and rely on the virus itself to kill the infected cells. Unfortunately, some cells infected by the virus memory T-cells are not killed by the virus.

T-cells are a type of lymphocyte, or white blood cell, produced by the thymus gland, that actively participates in the bodys immune response. Memory T-cells can live for years, or even decades, providing life-long immunity to previously encountered diseases. They can form "quiescent" infections, which last for years, and cause HIV to rebound whenever a patient stops treatment.

During a decade-long study, the researchers discovered that not all memory T-cells are alike. A subgroup of memory T-cells, called "Stem-Cell Memory T-cells" (Tscm), are different, particularly in their ability to produce daughter cells.

The researchers were able to show that the HIV-infected Tscm cells in patients on HIV therapy decayed more slowly than any other type of T-cell. As a result, after 10 years of therapy, the Tscm cells represented 24 percent of the total HIV infected cell population, despite being only 1 percent of the total T-cell population.

This finding is significant, Zurakowski said, because it demonstrates that Tscm cells are the slowest-decaying portion of the HIV reservoir.

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T-Cell Finding Sheds Light on Why HIV Can Persist Despite Treatment

Study identifies population of stem-like cells where HIV persists in spite of treatment

By Stem Cell Treatment

Jan. 12, 2014 Although antiviral therapy against HIV suppresses viral replication and allows infected individuals to live relatively healthy lives for many years, the virus persists in the body, and replication resumes if treatment is interrupted. Now investigators from Massachusetts General Hospital (MGH) and the Ragon Institute of MGH, MIT and Harvard may have found where the virus hides -- in a small group of recently identified T cells with stem-cell-like properties.

"Most human cells are short lived, so it has been unclear how HIV manages to stick around for decades in spite of very effective antiviral treatment," says Mathias Lichterfeld, MD, of the MGH Infectious Disease Division, corresponding author of the report receiving advance online publication in Nature Medicine. "This question led to the hypothesis that HIV might infect stem cells -- the most long-lasting cells in the body -- but traditional organ-specific stem cells, even those that give rise to all immune and blood cells, are resistant to HIV infection. We have discovered that a new group of T cells, called T memory stem cells, are susceptible to HIV and likely represent the longest lasting cellular niche for the virus."

HIV has such a devastating impact on the human immune system because it infects the CD4-positive T cells that normally direct and support the infection-fighting activities of other immune cells. Several subtypes of CD4 T cells have different functions; and all are capable of being infected by HIV, although antiviral treatment keeps the virus in those cells from replicating. Most of these CD4 T cells are short-lived and die relatively soon. What is distinct about CD4 T memory stem cells is their ability to live for decades, while giving rise to several subgroups of T cells. Therefore, HIV-infected T memory stem cells could continuously regenerate new HIV-infected cells, fueling the fire of HIV persistence in the human body.

The MGH/Ragon team found that T memory stem cells express both CD4 and CCR5 -- the receptor proteins used by HIV to enter cells -- suggesting that these long-lived cells could be the long-sought HIV reservoir. They then found that these cells can be readily infected with HIV, which was unexpected since traditional stem cells resist HIV infection. Importantly, the investigators found that levels of HIV DNA in patients receiving long-term antiviral treatment were highest in T memory stem cells.

Testing blood samples that had been taken from patients soon after initial infection and several years later revealed that the viral sequences found in T memory stem cells after 6 to 10 years of treatment were similar to those found in circulating T cells soon after infection, indicating that HIV had persisted relatively unchanged in T memory stem cells. In addition, the amount of HIV DNA in these cells remained relatively stable over time, even after long-term treatment caused viral levels to drop in other T cell subsets.

"Our findings suggest that novel, specific interventions will have to be designed to target HIV-infected T memory stem cells," says Lichterfeld, an assistant professor of Medicine at Harvard Medical School. "Methods of inhibiting stem cell pathways are being studied to eliminate cancer stem cells -- persistent cells that are responsible for tumor recurrence after conventional treatments kill proliferating tumor cells. We are now investigating whether any of the drugs that target cancer stem cells might be effective against HIV-infected T memory stem cells.

"Identifying the reservoirs for HIV persistence is a critical step toward developing interventions that could induce a long-term remission without the need for antiviral medication, or possibly eliminate the virus entirely," Lichterfeld adds. "Although a real cure for HIV has been elusive, it is not impossible."

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Study identifies population of stem-like cells where HIV persists in spite of treatment

New Treatment For Blood Diseases Using Artificial Bone Marrow

By Stem Cell Treatment

January 12, 2014

Image Caption: Scanning electron microscopy of stem cells (yellow / green) in a scaffold structure (blue) serving as a basis for the artificial bone marrow. Credit: C. Lee-Thedieck/KIT

Rebekah Eliason for redOrbit.com Your Universe Online

An exciting breakthrough is offering hope for the treatment of blood diseases such as leukemia using artificial bone marrow.

Specialized cells, known as hematopoietic stem cells, located within bone marrow, continuously replace and supply new blood cells such as red blood cells and white blood cells. Traditionally a blood disease like leukemia is treated with bone marrow transplants that supply the patient with new hematopoietic stem cells. Researchers have now discovered a way to artificially reproduce hematopoietic stem cells.

Since not every leukemia patient can find a suitable transplant, there is a need for other forms of treatment. The lack of appropriate transplants could be solved by artificial reproduction of hematopoietic stem cells. Previously, reproduction of the cells has been impossible due to their inability to survive anywhere but in their natural environment. Hematopoietic stem cells are found in a special niche of the bone marrow. If the cells reside out of the bone marrow, the specialized properties are modified. Consequently, to effectively reproduce the cells, the stem cell niche environment must also be created.

In the microscopic environment of the stem cell niche, there are several specific properties of importance. Areas in the bone that house the stem cells are extremely porous like a sponge. Making things even more complex, the spongy tissue is also home to other cell types which exchange signal substances with the stem cells. Also, the space among the cells creates an environment ensuring stability along with a place for the cells to anchor. Furthermore, the stem cell niche supplies the cells with nutrients and oxygen.

Dr. Cornelia Lee-Thedieck is head of the Young Investigators Group Stem Cell-Material Interactions, which consists of scientitsts from the KIT Institute of Functional Interfaces (IFG), the Max Planck Institute for Intelligent Systems, Stuttgart and Tbingen University. The team was successful at artificially reproducing major properties of bone marrow at the laboratory.

Using synthetic polymers, the researchers were able to create a porous structure that simulated the spongy environment of the blood-forming bone marrow. Also, they were able to add protein building blocks which are similar to those found naturally in the environment of the bone marrow that enable cells to anchor. Finally, they added the other types of cells needed for exchanging signaling substances.

After the artificial bone marrow was created, the scientists placed hematopoietic stem cells that had been isolated from cord blood into it. For several days the cells were bred. Various analytical methods were then used to determine that cells were able to reproduce in the artificial bone marrow. When compared with standard cell cultivation methods, a larger number of stem cells in the artificial bone marrow retained their specific properties.

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New Treatment For Blood Diseases Using Artificial Bone Marrow

Start of stem-cell study offers hope to patients with spinal-cord injuries

By Stem Cell Doctors

CTVNews.ca Staff Published Friday, January 10, 2014 4:33PM EST Last Updated Friday, January 10, 2014 11:42PM EST

A team of doctors at the University of Calgary has, for the first time in North America, successfully performed a stem cell transplant in a spinal cord injury patient, a procedure that could offer a glimmer of hope to patients whose injuries have long been considered untreatable.

The doctors injected the neural stem cells into the spine of a 29-year-old paraplegic, who will now be monitored to determine whether implanting those cells is safe.

Later studies will look at whether it is possible to regenerate new tissue and repair the mans injury.

That is the goal, a cure, the University of Calgarys Dr. Steven Casha, who performed the procedure on Wednesday, told CTV News.

Stem cells have the potential to recreate lost tissue, he added, although that remains to be proven in humans with spinal cord injuries. The answer, he said, is a long way away.

The transplant is part of an ongoing clinical trial being conducted by StemCells Inc., which harvested the stem cells from the nervous system of a fetus. The company holds a patent on the cells.

Data from three patients in Europe who have already undergone a transplant suggests the procedure is safe.

We have not been seeing significant complications or adverse eventsand there have been a couple of patients who havemade very small gains in functionthat appear to be hopeful and that is very interesting, Dr. Michael Fehlings, head of the spinal program at Toronto Western Hospital and the lead investigator for the trial at the University of Toronto, told CTV.

Fehlings cautioned that the results are very preliminary.

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Start of stem-cell study offers hope to patients with spinal-cord injuries

Artificial Bone Marrow Created By German Scientists, Could Be Used To Treat Leukemia Someday

By Stem Cell Doctors

Bone marrow nurtures both red blood cells and white blood cells, with healthy people producing more than 500 billion red- and-white blood cells every day. But when bone marrow is damaged by a disease like leukemia, or by radiation or chemotherapy drugs, the supply of blood cells drops, leaving a person at risk for fatal infections.

Leukemia and other types of bone-marrow diseases are often treated by transplanting healthy hematopoietic stem cells, which can develop into various kinds of blood cells, from another person. The donor cells can be taken from another persons bone marrow or bloodstream, or from preserved umbilical cords and placentas. But finding a matching donor can be difficult, and the amount of stem cells harvested from the donor may not always be enough to meet the needs of the patient.

One thing that doctors want to be able to do is to find a way to cultivate a bumper crop of stem cells. But blood stem cells thrive in a very specific environment inside bone marrow. And bone marrow has a very complex architecture, like a tiny sponge that contains many sizes of pores, and special docking proteins for stem cells.

"We assume that stem cells [do] not only notice the chemical composition of their surroundings., Karlsruhe Institute of Technology researcher and co-author of the study Cornelia Lee-Thedieck told German broadcaster Deutsche Welle. They can probably also feel if their environment is soft or hard, rough or smooth.

Lee-Thedieck and colleagues used a simple, porous polymer to mimic a sponge-like structure for the base of their artificial bone marrow. They added proteins similar to ones found in bone marrow to act as docking points for the blood stem cells, and added other cells to help ferry necessary molecular messages and materials back and forth.

When hematopoietic stem cells from cord blood were introduced to the artificial environment, they thrived much better than in standard 2-dimensional cell-culture systems. But the authors guess that it will be at least another 15 years before most patients will be able to benefit from this invention.

"Producing artificial bone marrow for culturing and multiplying blood stem cells is a potentially interesting application," Martin Bornhuser, a researcher from the University Hospital Dresden unaffiliated with the current paper, told DW. "It would make it possible to generate a sufficient number of stem cells from a small amount to transplant into an adult patient.

SOURCE: Raic et al. Biomimetic macroporous PEG hydrogels as 3D scaffolds for the multiplication of human hematopoietic stem and progenitor cells. Biomaterials 35: 929-940, January 2014.

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Artificial Bone Marrow Created By German Scientists, Could Be Used To Treat Leukemia Someday

Medical Board Suspends Oregon Doctor For Stem Cell Treatments …

By Stem Cell Medicine

Oregon Medical Board has issued a rare emergency suspension of a Eugene physicians license after the doctor conducted experimental stem cell treatments on patients.

The board considers Dr. Kenneth Welkers medical practice an immediate danger to the public.

Welker can appeal the suspension, issued Thursday. He did not return calls from The Associated Press on Friday, nor did the clinic at which hes employed, Oregon Optimal Health.

According to his online biography, Welker is a trained surgeon who quit his practice to pursue alternative medicine in 2007.

In May 2013, the boards suspension order says Welker injected processed stem cells into the spine of a 62-year-old woman, and was confused when she began to sweat and feel tingling in her extremities.

Stem cells, unlike other cells in the body, have two distinct characteristics. They can renew themselves through cell division, and they are not specialized in the way that muscle cells or brain cells are. Under certain conditions, they can be induced to transform into organ- or tissue-specific cells.

In 1998, researchers discovered how to derive stem cells from human embryos, and in 2006, they determined how to induce some specialized adult cells to take on the genetic characteristics of stem cells. These are called induced pluripotent stem cells, or iPSC.

iPSC have long been used to treat cancers such as leukemia and lymphoma its what doctors are using when they do bone marrow transplants. The cells are being studied for everything from heart disease to diabetes, but its too soon to know if these approaches are safe or effective.

Advocates of alternative medicine have heaped praise on the possibility of using iPSC to treat a variety of maladies. Texas Gov. Rick Perry, for instance, had stem cells taken from fat in his own body, grown in a lab and then injected into his back and his bloodstream during a 2011 operation to fuse part of his spine.

But scientists have questioned the safety and wisdom of Perrys treatment, especially because it was not part of a clinical trial in which unproven therapies are tested in a way that helps protect patients and advances medical knowledge.

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