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Major Advancement Towards Creating Patient-Specific and Disease-Specific Stem Cells For Therapeutic Use

Stemagen, a privately held embryonic stem cell research company, announced today it has become the first in the world to create, and meticulously document, a cloned human embryo using somatic cell nuclear transfer (SCNT).

Stemagen CEO Samuel H. Wood, M.D., Ph.D., a co-author of the publication and a donor of the cells from which the embryos were cloned, terms this achievement “a critical milestone in the development of patient-specific embryonic stem cells for human therapeutic use, potentially including developing treatments for Parkinson’s, Alzheimer’s and other degenerative diseases.” Stemagen’s research is exhaustively detailed in a paper published in today’s issue of the highly regarded peer-reviewed scientific journal Stem Cells.

“This is not merely a technical improvement on previous research in this area,” said Andrew French, Ph.D., lead author on the paper, “Development of Human Cloned Blastocysts Following Somatic Cell Nuclear Transfer (SCNT) with Adult Fibroblasts.”

“No other scientific group has documented the cloning of an adult human cell, much less been able to grow it to the blastocyst stage, the stage at which it is the adult donor cell that is driving embryonic development, the stage that yields the cells (the inner cell mass) from which embryonic stem cell lines are made,” said French, who is Stemagen’s Chief Scientific Officer.
Five blastocysts were developed from 25 donated mature oocytes. Three were confirmed to be clones based on DNA fingerprinting demonstrating the presence of the skin cell donor DNA in the blastocyst, while one was further confirmed to be a clone by an additional mitochondrial DNA (mtDNA) analysis which revealed the presence of oocyte donor mtDNA without any oocyte donor nuclear DNA. For technical reasons, the genetic material in the remaining two blastocysts did not amplify to the extent required for analysis, and so while it is likely they were clones, the evidence required to claim that with certainty was not present. Thus, in this study, cloned blastocysts were successfully created from approximately 10% of all mature donated oocytes, an unexpectedly high rate given past research in this field.

The oocytes used in this study were donated, without compensation, by egg donors and intended parents undergoing egg donation cycles for reproductive purposes at the Reproductive Sciences Center in La Jolla, a leading fertility center specializing in egg donation and other advanced assisted reproductive technologies.

“As important as stem cell research is, all of us involved in this study realized that our overriding responsibility was to the intended parents who entrusted us with their dream of having a child,” said Catharine Adams, Ph.D., a co-author on the paper and the laboratory director for Reproductive Sciences Center. “We in the IVF laboratory felt comfortable in this collaboration because we have consistently achieved pregnancy rates of greater than 80% from these types of high quality egg donors. In this study, all the intended parents were successful in achieving a pregnancy.”

Stemagen and the Reproductive Sciences Center worked closely, over an extended period of time, with a leading independent Institutional Review Board (IRB) to develop procedures ensuring that all parties received comprehensive informed consent and that procedures were in place to protect their confidentiality in the process. All research procedures, including the culturing of the skin cells (fibroblasts) were performed under clinical laboratory conditions in close cooperation with the Assisted Reproductive Technologies (ART) Laboratory of the Reproductive Sciences Center, directed by Catharine Adams, Ph.D. French notes, “An important reason for the success of our SCNT procedures depended on the close coordination between our laboratory personnel and fertility center laboratory staff. Timing is a critical element in maximizing the probability of success in this type of procedure.”

Wood points out that this research was exhaustively scrutinized by some of the world’s most respected scientists and underwent an exceptionally rigorous process of verification, “This achievement was so critical to our field, we felt we should spare no effort in the process of establishing the validity of our work.”

DNA fingerprinting is the scientifically accepted method for determining if an embryo is a true clone. According to French, “All samples were subjected to this type of analysis to determine their true genetic makeup.”

For that, the company turned to Genesis Genetics, a recognized worldwide leader in the field of reproductive embryonic analysis.

Company founder and CEO, Mark Hughes, M.D., Ph.D., said “We were proud to collaborate with Stemagen in this important accomplishment. As the leading provider of genetic diagnosis of human embryos, it was important for an independent company like Genesis Genetics to be involved in the verification of this achievement.

Stemagen, Inc., is dedicated to the production of patient-specific embryonic stem cells for therapeutic use through SCNT and “uniparental” embryonic stem cells technology.

http://www.stemagen.com/17jan08.htm

How Important Is the Latest Cloning Feat?

Scientists at Stemagen, a small biotechnology company in La Jolla, CA, reported yesterday that they have for the first time generated cloned human blastocysts--early-stage embryos--from adult skin cells. This is the first step in generating stem cell lines matched to individuals, which are crucial for creating new cellular models of disease and potentially important for future tissue replacement therapies. (See "Next Steps for Stem Cells" and "The Real Stem Cell Hope".) The new findings also confirm that access to fresh eggs from healthy young donors is a key part of successful cloning. Lack of access to human eggs has been the major barrier in the field. (See "Human Therapeutic Cloning at a Standstill".)

Cloned blastocysts have been generated before, but from embryonic stem cells rather than from adult cells. Scientists theorize that embryonic stem cells are easier to turn into blastocysts because of their earlier developmental stage.

Experts in the field have had a mixed reaction to the new work. "It's a nice achievement, but in my view, they haven't crossed the bar," says Evan Snyder, director of the Stem Cells and Regenerative Medicine Program at the Burnham Institute in La Jolla. "The real test will be, can you generate cell lines that are stable and self-renewing and normal?" Others applaud the confirmation of the feasibility of human cloning. "The fact that it can be done is important," says Jeanne Loring, a stem cell scientist at the Scripps Research Institute in La Jolla. "It wipes away that blot on our scientific integrity," she says, referring to a massive fraud unveiled in 2005 in which South Korean scientist Woo Suk Hwang claimed to have generated stem cell lines from cloned human embryos. (See "Stem Cells Reborn".)

To clone an embryo, a process also called nuclear transfer, scientists first strip an egg of its genetic material. Then they insert DNA from an adult cell, such as a skin cell, into the egg. Through an unknown process, the egg turns back the clock on the adult DNA and begins to develop as a normally fertilized egg would. From the embryo, researchers could theoretically collect a specialized ball of cells that can be coaxed to turn into stem cells. So far, however, no one has successfully performed this feat.

Stemagen, a relatively unknown player in the field, probably owes its success to access to human eggs through a close association with a local fertility clinic. (The company was founded by a fertility specialist at the Reproductive Sciences Center in La Jolla.) "We were able to get access to high-quality oocytes and have them in the incubator within one to two hours," says Andrew French, Stemagen's chief scientific officer.

Egg donors and the intended parents gave eggs in excess of those needed for in vitro fertilization to the Stemagen scientists for research. Regulations in many states prohibit compensation for donated eggs for ethical reasons, a requirement that has slowed other cloning efforts.


Starting with 25 fresh oocytes, French and colleagues generated five blastocysts--five- to six-day-old embryos consisting of 30 to 70 cells. Rather than attempting to generate stem cell lines from the embryos, the researchers sent them to an independent company for genetic confirmation of their results. "They showed we had completely removed the DNA from the egg donor and replaced it with DNA from the skin-cell donor," says French. One blastocyst was confirmed as a clone via two DNA-fingerprinting methods, while genetic analysis of two others indicated the likelihood that they were clones.

The next crucial step will be generating stem cell lines from cloned embryos, which many stem cell scientists speculate will be the most challenging step. "That's likely where Hwang failed," says Synder.

French and colleagues are planning such experiments, with results potentially in the next eight to twelve months. "The quality of our blastocysts improved with each experiment," says French. Based on the success rate of previous attempts to make stem cells from regular embryos, he estimates that Stemagen will be able to generate a stem cell line from between five and ten cloned embryos and report the results in the next year. The company aims to sell or license the lines to pharmaceutical companies and others who would use them to test new drugs or develop new therapies.

While human therapeutic cloning has always been an ethically contentious area of research--partly because it requires the creation and destruction of human embryos--it has recently come under greater fire. After the announcement of new techniques for reprogramming adult cells so that they turn into stem cells without first forming embryos, some opponents called for a halt on embryonic-stem-cell research. (See "Stem Cells without the Embryos".)

However, researchers in the field emphasize the need to pursue all reprogramming techniques. "Even though there are other techniques to reprogram a cell that have gotten a lot of press, we still don't know how those compare with the reprogramming you actually see with nuclear transfer," says Snyder. "My feeling is, if we understand nuclear transfer better, we will be able to do the other kind of reprogramming more efficiently."

http://www.technologyreview.com/Biotech/20088/

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DNA Fingerprints Predict Brain Disorders

Predicting brain disorders such as Parkinson's disease and amyotrophic lateral sclerosis (ALS) may be possible by identifying certain DNA fingerprints, a recent Mayo Clinic study suggests.
The researchers analyzed the genetic data of people with ALS (also known as Lou Gehrig's disease) and Parkinson's, and those without neurological disorders. They pinpointed several gene variations that predicted who was at higher risk for developing the disorders, according to the study published online in PLoS One.

The variations were found by studying the axon guidance pathway, a complex collection of chemical signals that wire the brain during fetal development, and maintain and repair brain circuits throughout a person's life.

Researchers found many variations within pathway genes common to ALS and Parkinson's. However, they also identified several that collectively indicate people at high risk (2,000 times greater than the general population) for ALS. And they pinpointed several gene variations that collectively predicted people at high risk (nearly 400 times greater than the general population) for Parkinson's disease.

"In persons at high risk, we may be able to prevent the diseases or slow or halt their progression by developing drugs that target the same disease pathways. For ALS and Parkinson's disease, our study is a major step in these directions," principal investigator and Mayo Clinic neurologist Dr. Demetrius Maraganore said in a prepared statement.

ALS is a progressive neurodegenerative disease that leads to loss of muscle control and use, and eventually death. As many as 30,000 Americans have the disease at any given time, according to the ALS Association. Parkinson's disease, a brain disorder that affects 1.5 million Americans, causes tremors, slowness of movement, and body stiffness, according to the National Parkinson Foundation. No cure exists for either disease.

http://www.businessweek.com/lifestyle/content/healthday/611747.html?chan=search