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    AUA 2013: Early steps taken in using stem cells for infertility

    Wake Forest University researchers have conducted early, pre-clinical work that they say supports further research on the use of spermatogonial stem cell (SSC) transplantation as a potential treatment for male infertility.

    SSC autotransplantation would play a particularly important role in pediatric cancer patients undergoing chemotherapy or radiation therapy, in whom sterility cannot be prevented and who have no means available to preserve fertility, said John D. Jackson, PhD, of the Wake Forest University School of Medicine and the Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC.

    The research is being led by Anthony Atala, MD, and Hooman Sadri-Ardekani, PhD. Dr. Sadri-Ardekani has developed a culture technique aimed at propogating stem cells, and the researchers are in the process of characterizing those cells using a panel of three phenotype markers.

    “The only way to truly identify a stem cell is to go back into a human and show that you get repopulation of cells. But you can use these other assays as surrogates,” Dr. Johnson said.

    In the current study, adult cadaveric human testicular tissues were received from three individuals, and the tissues were immediately cryopreserved. After thawing and enzymatic digestion, the testicular cells were seeded. The presence of spermatogonial cells and their quantity were evaluated by germ line stem cell (GSC) formation, reverse transcriptase (RT) PCR, and flow cytometry.

    The first GSC cluster formation was seen after 2 weeks of culture, Dr. Johnson reported. RT PCR for PLZF (ZBTB16), UCHL1 (PGP 9.5), FGFR3, and CD9 as markers for undifferentiated spermatogonial cells (including SSCs) confirmed the presence of these cells during the culture period. Flow cytometric analyses showed various percentages (0.7%-6%) of HLA-ABC-negative/FGFR3-positive cells (spermatogonial cells) in different passages during the culture period.

    “We’re in the very early stages. Clinical trials haven’t been started. We’re collecting tissue now under an approved IRB,” Dr. Johnson said.

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