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Newswise MADISON, Wis. As stem cells continue their gradual transition from the lab to the clinic, a research group at the University of Wisconsin-Madison has discovered a new way to make large concentrations of skeletal muscle cells and muscle progenitors from human stem cells.

The new method, described in the journal Stem Cells Translational Medicine, could be used to generate large numbers of muscle cells and muscle progenitors directly from human pluripotent stem cells. These stem cells, such as embryonic (ES) or induced pluripotent stem (iPS) cells, can be made into virtually any adult cell in the body.

Adapting a method previously used to make brain cells, Masatoshi Suzuki, an assistant professor of comparative biosciences in the School of Veterinary Medicine, has directed those universal stem cells to become both adult muscle cells and muscle progenitors.

Importantly, the new technique grows the pluripotent stem cells as floating spheres in high concentrations of two growth factors, fibroblast growth factor-2 and epidermal growth factor. These growth factors "urge" the stem cells to become muscle cells.

"Researchers have been looking for an easy way to efficiently differentiate stem cells into muscle cells that would be allowable in the clinic," says Suzuki. The novelty of this technique is that it generates a larger number of muscle stem cells without using genetic modification, which is required by existing methods for making muscle cells.

"Many other protocols have been used to enhance the number of cells that go to a muscle fate," says co-author Jonathan Van Dyke, a post-doctoral fellow in Suzukis laboratory. "But what's exciting about the new protocol is that we avoid some techniques that would prohibit clinical applications. We think this new method has great promise for alleviating human suffering."

Last year, Suzuki demonstrated that transplants of another type of human stem cells somewhat improved survival and muscle function in rats that model amyotrophic lateral sclerosis (ALS). Also known as Lou Gehrig's disease, ALS destroys nerves and causes a loss of muscle control. The muscle progenitors generated with Suzukis new method could potentially play a similar role but with enhanced effect.

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A New Way to Make Muscle Cells From Human Stem Cells

London, March 21 (IANS): In a major breakthrough, a team of scientists has developed a method to generate human induced pluripotent stem cells (hiPSCs) from just a drop of finger-pricked blood.

As hiPSCs exhibit properties remarkably similar to human embryonic stem cells, they are invaluable resources for scientific research.

Earlier methods to generate hiPSCs generally required large quantities of blood.

The new method developed by scientists at Institute of Molecular and Cell Biology (IMCB), Singapore also enables donors to collect their own blood samples which they can then send to a laboratory for further processing.

The do it yourself (DIY) finger-prick technique is the worlds first to use only a drop of finger-pricked blood to yield hiPSCs with high efficiency.

It all began when we wondered if we could reduce the volume of blood used for (genetic) reprogramming. We then tested if donors could collect their own blood sample in a normal room environment and store it, said Loh Yuin Han Jonathan, principal investigator at IMCB.

Our finger-prick technique, in fact, utilised less than a drop of finger-pricked blood. The remaining blood could even be used for DNA sequencing and other blood tests, Jonathan said.

The accessibility of the new technique is further enhanced with a DIY sample collection approach.

The blood sample remains stable for 48 hours and can be expanded for 12 days in culture, which therefore extends the finger-prick technique to a wide range of geographical regions for recruitment of donors with varied ethnicities, genotypes and diseases.

By integrating it with the hiPSC bank initiatives, the finger-prick technique has paved the way for establishing diverse and fully characterised hiPSC banking for stem cell research.

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Stem cells generated from just blood!

For the first time, scientists have succeeded in coaxing laboratory cultures of human stem cells to develop into the specialized, unique cells needed to repair a patient's defective or diseased bladder.

The breakthrough, developed at the UC Davis Institute for Regenerative Cures and published today in the scientific journal Stem Cells Translational Medicine, is significant because it provides a pathway to regenerate replacement bladder tissue for patients whose bladders are too small or do not function properly, such as children with spina bifida and adults with spinal cord injuries or bladder cancer.

"Our goal is to use human stem cells to regenerate tissue in the lab that can be transplanted into patients to augment or replace their malfunctioning bladders," said Eric Kurzrock, professor and chief of the division of pediatric urologic surgery at UC Davis Children's Hospital and lead scientist of the study, which is titled "Induction of Human Embryonic and Induced Pluripotent Stem Cells into Urothelium."

To develop the bladder cells, Kurzrock and his UC Davis colleagues investigated two categories of human stem cells. In their key experiments, they used induced pluripotent stem cells (iPS cells), which were derived from lab cultures of human skin cells and umbilical blood cells that had been genetically reprogrammed to convert to an embryonic stem cell-like state.

If additional research demonstrates that grafts of bladder tissue grown from human stem cells will be safe and effective for patient care, Kurzrock said that the source of the grafts would be iPS cells derived from a patient's own skin or umbilical cord blood cells. This type of tissue would be optimal, he said, because it lowers the risk of immunological rejection that typifies most transplants.

In their investigation, Kurzrock and his colleagues developed a protocol to prod the pluripotent cells into becoming bladder cells. Their procedure was efficient and, most importantly, the cells proliferated over a long period of time -- a critical element in any tissue engineering application.

"What's exciting about this discovery is that it also opens up an array of opportunities using pluripotent cells," said Jan Nolta, professor and director of the UC Davis Stem Cell program and a co-author on the new study. "When we can reliably direct and differentiate pluripotent stem cells, we have more options to develop new and effective regenerative medicine therapies. The protocols we used to create bladder tissue also provide insight into other types of tissue regeneration."

UC Davis researchers first used human embryonic stem cells obtained from the National Institutes of Health's repository of human stem cells. Embryonic stem cells can become any cell type in the body (i.e., they are pluripotent), and the team successfully coaxed these embryonic stem cells into bladder cells. They then used the same protocol to coax iPS cells made from skin and umbilical cord blood into bladder cells, called urothelium, that line the inside of the bladder. The cells expressed a very unique protein and marker of bladder cells called uroplakin, which makes the bladder impermeable to toxins in the urine.

The UC Davis researchers adjusted the culture system in which the stem cells were developing to encourage the cells to proliferate, differentiate and express the bladder protein without depending upon signals from other human cells, said Kurzrock. In future research, Kurzrock and his colleagues plan to modify the laboratory cultures so that they will not need animal and human products, which will allow use of the cells in patients.

Kurzrock's primary focus as a physician is with children suffering from spina bifida and other pediatric congenital disorders. Currently, when he surgically reconstructs a child's defective bladder, he must use a segment of their own intestine. Because the function of intestine, which absorbs food, is almost the opposite of bladder, bladder reconstruction with intestinal tissue may lead to serious complications, including urinary stone formation, electrolyte abnormalities and cancer. Developing a stem cell alternative not only will be less invasive, but should prove to be more effective, too, he said.

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Stem cell findings may offer answers for some bladder defects, disease

PUBLIC RELEASE DATE:

21-Mar-2014

Contact: Charles Casey charles.casey@ucdmc.ucdavis.edu 916-734-9048 University of California - Davis Health System

(SACRAMENTO, Calif.) For the first time, scientists have succeeded in coaxing laboratory cultures of human stem cells to develop into the specialized, unique cells needed to repair a patient's defective or diseased bladder.

The breakthrough, developed at the UC Davis Institute for Regenerative Cures and published today in the scientific journal Stem Cells Translational Medicine, is significant because it provides a pathway to regenerate replacement bladder tissue for patients whose bladders are too small or do not function properly, such as children with spina bifida and adults with spinal cord injuries or bladder cancer.

"Our goal is to use human stem cells to regenerate tissue in the lab that can be transplanted into patients to augment or replace their malfunctioning bladders," said Eric Kurzrock, professor and chief of the division of pediatric urologic surgery at UC Davis Children's Hospital and lead scientist of the study, which is titled "Induction of Human Embryonic and Induced Pluripotent Stem Cells into Urothelium."

To develop the bladder cells, Kurzrock and his UC Davis colleagues investigated two categories of human stem cells. In their key experiments, they used induced pluripotent stem cells (iPS cells), which were derived from lab cultures of human skin cells and umbilical blood cells that had been genetically reprogrammed to convert to an embryonic stem cell-like state.

If additional research demonstrates that grafts of bladder tissue grown from human stem cells will be safe and effective for patient care, Kurzrock said that the source of the grafts would be iPS cells derived from a patient's own skin or umbilical cord blood cells. This type of tissue would be optimal, he said, because it lowers the risk of immunological rejection that typifies most transplants.

In their investigation, Kurzrock and his colleagues developed a protocol to prod the pluripotent cells into becoming bladder cells. Their procedure was efficient and, most importantly, the cells proliferated over a long period of time a critical element in any tissue engineering application.

"What's exciting about this discovery is that it also opens up an array of opportunities using pluripotent cells," said Jan Nolta, professor and director of the UC Davis Stem Cell program and a co-author on the new study. "When we can reliably direct and differentiate pluripotent stem cells, we have more options to develop new and effective regenerative medicine therapies. The protocols we used to create bladder tissue also provide insight into other types of tissue regeneration."

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Stem cell findings may offer answers for some bladder defects and disease

1 hour ago by Michael Mccarthy Julie Mathieu (left), Hannele Ruohola-Baker, and Zsuzsa Agoston go over laboratory research results.

Proteins that regulate energy metabolism are essential for stem cell formation, University of Washington researchers find.

Two proteins that control how cells metabolize glucose play a key role in the formation of human stem cells, UW researchers report.

The findings advance scientists' understanding of stem cell development but also suggest that the proteins, which also play a role in the process that transforms normal cells into cancer stem cells, might also be targets for new cancer therapies, the researchers write.

The findings appear online in the journal Cell Stem Cell. The paper's lead authors are Julie Mathieu, a post-doctoral fellow at the UW and Wenyu Zhou who was a graduate student at UW and is now a postdoctoral scholar at Stanford University, Department of Genetics. Dr. Hannele Ruohola-Baker, UW professor of biochemistry, is the paper's senior author.

In the study, the researchers induced mature human tissue fibroblasts to revert to an earlier stem cell-like state by inserting genes for four proteins, a process called reprogramming.

These reprogrammed cells have the extraordinary ability to develop into any type of cell in the human body, a capacity called pluripotency, and it is hoped that induced-pluripotent stem cells will one day be able to be used to create new tissues and organs to repair and replace those damaged by injury and disease.

Researches have known for some time that during reprogramming, cells must go through a stage in which they shut down metabolic pathway that they use to generate energy from glucose that requires the presence of oxygen in mitochondria, the cell's powerhouse and shift over to another pathway, called the glycolytic pathway, that generates less energy but does not require the presence of oxygen.

This shift may take place because in nature, embryonic and tissue stem cells often must survive in low-oxygen, or hypoxic, conditions.

This transition to a glycolytic state is of particular interest to cancer researchers as well, since as normal cells are transformed into cancer cells, which in many ways resemble stem cells, they, too, go through a glycolytic phase.

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Proteins that control energy use necessary to form stem cells

Dallas, TX (PRWEB) March 20, 2014

The Repair Stem Cells Institute (RSCI) -- http://www.repairstemcells.org -- is pleased to announce that it will assist interested patients to take part in a patient-sponsored research study based in the United States for the treatment of Type2 Diabetes with adult stem cells. The study, which meets current FDA guidelines, will be conducted during April 2014.

The study is being conducted by the U.S. based company Bioheart which has assembled teams of doctors and specialists specially trained in stem cell treatments. Based on previous treatment of Type 2 diabetes with autologous (the patients own) stem cells, it is estimated that two-thirds of participants will experience a significant quality of life improvement and symptoms reduction.

Type 2 diabetes makes up about 90% of cases of diabetes. Rates of type 2 diabetes have increased markedly since 1960. Today there are approximately 50 million people suffering from the disease compared to 15 million in 1985.

In a recent interview, RSCI founder and Chairman Don Margolis stated, With stem cell treatment rapidly coming to the forefront of 21st Century medicine, we are pleased that Type 2 Diabetes is among the many chronic conditions that are treatable with adult stem cells rather than potentially risky surgery, dangerous transplants, and toxic drugs.

Eligibility

Patients suffering from Type 2 diabetes who are cancer-free can apply to participate.

What will happen?

The 4-part procedure will be done in a participating doctors office as a point-of-care out-patient.

1.Adipose Harvest: During a 3 to 5 hour visit to the doctors office, a mini-liposuction on your stomach will extract a small amount of tummy fat containing tens of millions of adipose stem cells. 2.Laboratory Processing: The extracted stem cells will be isolated, analyzed, cleaned and concentrated. 3.Stem Cell Implantation: Up to 60 million stem cells will be transplanted intravenously, usually into your arm. Because these are the patients own cells, the risk of rejection is non-existent. 4.Postoperative Care: Normally, patients can leave shortly after implantation. RSCI will check on your progress monthly by telephone for the first year after stem cells.

Continued here:
The Repair Stem Cells Institute Invites Participation in a Unique Study of a Stem Cell Treatment for Type 2 Diabetes

Tokyo: One of the authors of a study that was claimed to have discovered a simple way to make stem cells has said that he is no longer sure of his team's conclusions and called for the study to be retracted.

The study, laid out in two papers published by Nature in January, surprised scientists around the world by finding that a simple acid bath might turn cells in the body into multi-purpose stem cells. The new technique could be a quicker and easier source of stem cells than methods now in use, the authors said.

But on Monday Teruhiko Wakayama, a professor of developmental engineering at the University of Yamanashi and one of the study's co-authors, told NHK, Japan's public broadcaster, that a series of concerns raised in recent weeks by researchers around the world had shaken his belief in the study's findings.

"There are too many overall issues that I am not sure about. I am increasingly uncertain," Dr Wakayama told NHK.

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The new technique was developed by researchers at the Riken Centre for Developmental Biology in Kobe, Japan, and at Brigham and Women's Hospital and Harvard Medical School in Boston. Haruko Obokata, the 30-year-old lead writer of the study's two papers and a rising star in Japan's scientific community, has become an overnight celebrity here and a symbol of the rising stature of female scientists.

Still, some experts quickly expressed caution, saying more needed to be known about the new approach.

Caution turned to scepticism as researchers reported trouble in replicating the study's results. Some of the photos used in the study were then called into question, as was wording that was found to be similar to that in an article published by different researchers almost a decade ago. Those questions prompted both Riken and Nature to begin separate investigations into the study's integrity last month.

Riken has since released a more detailed description of procedures used in the study. But inconsistencies between those new procedures and the original papers only fuelled confusion and suspicion. Nature has said it is still investigating.

Dr Wakayama said that the numerous questions raised left the authors with little choice but to retract the paper. Verification by independent researchers might also shed light on what went wrong in the study, he said.

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Author of stem cell study calls for retraction

The DIY finger-prick technique opens door for extensive stem cell banking

1. Scientists at A*STARs Institute of Molecular and Cell Biology (IMCB) have developed a method to generate human induced pluripotent stem cells (hiPSCs) from a single drop of finger-pricked blood. The method also enables donors to collect their own blood samples, which they can then send to a laboratory for further processing. The easy access to blood samples using the new technique could potentially boost the recruitment of greater numbers and diversities of donors, and could lead to the establishment of large-scale hiPSC banks.

3. Current sample collection for reprogramming into hiPSCs include invasive measures such as collecting cells from the bone marrow or skin, which may put off many potential donors. Although hiPSCs may also be generated from blood cells, large quantities of blood are usually required. In the paper published online on the Stem Cell Translational Medicine journal, scientists at IMCB showed for the first time that single-drop volumes of blood are sufficient for reprogramming into hiPSCs. The finger-prick technique is the worlds first to use only a drop of finger-pricked blood to yield hiPSCs with high efficiency. A patent has been filed for the innovation.

4. The accessibility of the new technique is further enhanced with a DIY sample collection approach. Donors may collect their own finger-pricked blood, which they can then store and send it to a laboratory for reprogramming. The blood sample remains stable for 48 hours and can be expanded for 12 days in culture, which therefore extends the finger-prick technique to a wide range of geographical regions for recruitment of donors with varied ethnicities, genotypes and diseases.

5. By integrating it with the hiPSC bank initiatives, the finger-prick technique paves the way for establishing diverse and fully characterised hiPSC banking for stem cell research. The potential access to a wide range of hiPSCs could also replace the use of embryonic stem cells, which are less accessible. It could also facilitate the set-up of a small hiPSC bank in Singapore to study targeted local diseases.

6. Dr Loh Yuin Han Jonathan, Principal Investigator at IMCB and lead scientist for the finger-prick hiPSC technique, said, It all began when we wondered if we could reduce the volume of blood used for reprogramming. We then tested ifdonors could collect their own blood sample in a normal room environment and store it. Our finger-prick technique, in fact, utilised less than a drop of finger-pricked blood. The remaining blood could even be used for DNA sequencing and other blood tests.

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:: 20, Mar 2014 :: A*STAR SCIENTISTS CREATE STEM CELLS FROM A DROP OF BLOOD

Durham, NC (PRWEB) March 19, 2014

In a study just published in STEM CELLS Translational Medicine, a group of researchers have discovered what appears to be an easy way to collect large quantities of viable stem cells that can be banked for future regenerative medicine purposes all from the simple prick of a finger.

We show that a single drop of blood from a finger-prick sample is sufficient for performing cellular reprogramming, DNA sequencing and blood typing in parallel. Our strategy has the potential of facilitating the development of large-scale human iPSC banking worldwide, said Jonathan Yuin-Han Loh, Ph.D., of the Agency for Science, Technology and Research (A*STAR) in Singapore. He is principal investigator on the study that also included scientists from other Singapore facilities as well as those in the United States and Great Britain.

The medical world in general is excited about the potential of induced pluripotent stem cells (iPSCs) for studying diseases and for therapeutic regenerative medicine. Stem cells harvested from bone marrow and cord blood are highly amenable to reprogramming.

Some methods can result in negative side effects, and then you have bone-marrow harvesting, which is invasive, while cord blood is limited to individuals who have deposited their samples at birth, Dr. Loh explained. The large amount of blood needed to collect enough cells for reprogramming has also deterred many potential donors.

"We gradually reduced the starting volume of blood (collected using a needle) and confirmed that reprogramming can be achieved with as little as .25 milliliters, Hong-kee Tan, lead author on the study and a research officer in the Loh lab reported.

This then made the team wonder whether a DIY (do-it-yourself) approach to blood collection might work too.

To test this idea, we asked donors to prick their own fingers in a normal room environment and collect a single drop of blood sample into a tube, Tan said. The tube was placed on ice and delivered to the lab for reprogramming.

The cells were treated with a buffer at 12-, 24- or 48-hour increments and observed under the microscope for viability and signs of contamination. After 12 days of expansion in medium, the cells appeared healthy and were actively dividing. The team next tested what happened when they reprogrammed the cells and succeeded in forcing them to become mesodermal, endodermal and neural cells. They were even able to induce some into giving rise to rhythmically beating cardiomyocytes.

Interestingly, we did not observe any noticeable reduction in reprogramming efficiency between the freshly collected and the DIY finger-prick samples, Dr. Loh said. In summary, we derived healthy iPSCs from tiny volumes of venipuncture and a single drop finger-prick blood samples. We also report a high reprogramming yield of 100 to 600 colonies per milliliter of blood.

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DIY Finger Prick Yields Ample Stem Cells for Banking

PUBLIC RELEASE DATE:

18-Mar-2014

Contact: Anita Srikameswaran 412-578-9193 University of Pittsburgh Schools of the Health Sciences

PITTSBURGH, March 18, 2014 Stem cells derived from human muscle tissue were able to repair nerve damage and restore function in an animal model of sciatic nerve injury, according to researchers at the University of Pittsburgh School of Medicine. The findings, published online today in the Journal of Clinical Investigation, suggest that cell therapy of certain nerve diseases, such as multiple sclerosis, might one day be feasible.

To date, treatments for damage to peripheral nerves, which are the nerves outside the brain and spinal cord, have not been very successful, often leaving patients with impaired muscle control and sensation, pain and decreased function, said senior author Johnny Huard, Ph.D., professor of orthopaedic surgery, and Henry J. Mankin Chair in Orthopaedic Surgery Research, Pitt School of Medicine, and deputy director for cellular therapy, McGowan Institute for Regenerative Medicine.

"This study indicates that placing adult, human muscle-derived stem cells at the site of peripheral nerve injury can help heal the lesion," Dr. Huard said. "The stem cells were able to make non-neuronal support cells to promote regeneration of the damaged nerve fiber."

The researchers, led by Dr. Huard and Mitra Lavasani, Ph.D., first author and assistant professor of orthopaedic surgery, Pitt School of Medicine, cultured human muscle-derived stem/progenitor cells in a growth medium suitable for nerve cells. They found that, with prompting from specific nerve-growth factors, the stem cells could differentiate into neurons and glial support cells, including Schwann cells that form the myelin sheath around the axons of neurons to improve conduction of nerve impulses.

In mouse studies, the researchers injected human muscle-derived stem/progenitor cells into a quarter-inch defect they surgically created in the right sciatic nerve, which controls right leg movement. Six weeks later, the nerve had fully regenerated in stem-cell treated mice, while the untreated group had limited nerve regrowth and functionality. Twelve weeks later, treated mice were able to keep their treated and untreated legs balanced at the same level while being held vertically by their tails. When the treated mice ran through a special maze, analyses of their paw prints showed eventual restoration of gait. Treated and untreated mice experienced muscle atrophy, or loss, after nerve injury, but only the stem cell-treated animals had regained normal muscle mass by 72 weeks post-surgery.

"Even 12 weeks after the injury, the regenerated sciatic nerve looked and behaved like a normal nerve," Dr. Lavasani said. "This approach has great potential for not only acute nerve injury, but also conditions of chronic damage, such as diabetic neuropathy and multiple sclerosis."

Drs. Huard and Lavasani and the team are now trying to understand how the human muscle-derived stem/progenitor cells triggered injury repair, as well as developing delivery systems, such as gels, that could hold the cells in place at larger injury sites.

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Stem cells from muscle can repair nerve damage after injury, Pitt researchers show