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Novel Method To Grow Human Embryonic Stem Cells Developed
ScienceDaily (Aug. 20, 2008) - The majority of researchers working with 
human embryonic stem cells (hESCs) - cells which produce any type of 
specialized adult cells in the human body - use animal-based materials for 
culturing the cells. But because these materials are animal-based, they 
could transmit viruses and other pathogens to the hESCs, making the cells 
unsuitable for medical use.
Now, a stem-cell scientist at UC Riverside has devised a method of growing 
hESCs in the lab that uses no animal-derived materials - an important 
advance in the use of hESCs for future medical purposes.
Because of their tremendous potential, hESCs are considered promising 
sources for future cell therapy to treat diseases such as Parkinson's 
disease and diabetes mellitus.
Noboru Sato, an assistant professor of biochemistry, developed the new 
method, which is not only cleaner and easier to use than conventional 
methods of culturing hESCs but also results in hESCs whose pluripotency - 
the potential to differentiate into any of the specialized cells of the body 
such as neurons, cardiac muscles, and insulin-producing cells - is 
uncompromised.
Currently in labs worldwide, many researchers grow hESCs on Matrigel-coated 
culture plates, Matrigel being the trade name for a gelatinous extract, 
taken from mouse tumor cells, that contains extracellular matrices (ECMs), 
made up of special proteins. The Matrigel coating provides the scaffolding 
to which the hESCs first attach and then grow in undifferentiated colonies 
before differentiating into specialized cells.
"The development of animal-free coating methods for hESCs still remains a 
major challenge due to the complexity of ECMs and insufficient knowledge 
about how hESCs control cell-cell and cell-ECM interactions," explained 
Sato, who led the research project.
His lab identified a specific signaling pathway, called Rho-Rock, which the 
hESCs use during colony formation and which plays an important role in 
physical interactions between hESCs. When the researchers blocked the 
pathway, they found, as expected, that the normal colony formation of hESCs 
was considerably impaired. They also found that the hESCs maintained their 
pluripotency.
"Until now, it was generally assumed that the hESC colony formation was 
pivotal for maintaining pluripotency," Sato said. "But we show that 
pluripotency can be retained independent of close cell-cell contact."
Prue Talbot, the director of UCR's Stem Cell Center of which Sato is a 
member, noted that Sato's discovery could affect the way embryonic stem 
cells are grown in the future.
"His work is certainly an important step forward in both understanding 
signal transduction pathways in stem cells and in the development of an 
improved methodology for culturing stem cells," she said.
In the study, Sato's group extensively screened various types of scaffold 
materials in combination with Y27632, a chemical compound that blocks the 
Rho-Rock pathway, and found that the Matrigel coating could be replaced with 
"poly-D-lysine," a chemically synthesized ECM. The major advantages of 
poly-D-lysine over Matrigel are that poly-D-lysine is completely 
animal-free, easy to handle, and its quality is consistent.
"We found that the growth of the hESCs under this novel culture condition 
was almost identical to the growth of hESCs on Matrigel-coated culture 
plates, with no compromise in pluripotency," Sato said.
Having started his career as a physician in Japan, Sato began researching 
stem cell biology as a research fellow at The Rockefeller University, NY, 
one of the foremost research centers in the world. He accepted a faculty 
position in the Department of Biochemistry at UCR in 2006. He was joined in 
the research project by Nicole Harb of UCR and Trevor K. Archer of the 
National Institute of Environmental Health Sciences (NIEHS), NC.
The research was a collaboration between UCR and NIEHS, and funded by UCR 
start-up funds to Sato and a grant to Archer from the National Institutes of 
Health.
"Our research goal is to understand the basic mechanisms underlying unique 
biological functions of pluripotent stem cells, and to translate the 
obtained knowledge into future medical applications," Sato said.
His group is now focusing on applying his technique to the latest stem cell 
technology, "induced pluripotent stem (iPS) cells," which are pluripotent 
stem cells artificially derived from adult cells without using embryos. "Our 
next step is to produce new animal-free iPS cell lines," Sato said.
UCR's Office of Technology Commercialization has applied for a patent on 
Sato's discovery and is looking for industrial partners interested in 
further developing it.

Rayilyn Brown
Director AZNPF
Arizona Chapter National Parkinson Foundation
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