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BSP-13
(P)
ELUCIDATION OF REGULATORY MECHANISMS DURING DIFFERENTIATION OF
SPERMATOGONIAL STEM CELLS VIA ANDROGEN ACTION
Kentaro MIZUNO, Yutaro HAYASHI, Hideyuki KAMISAWA, Yoshinobu MORITOKI, Hidenori NISHIO, Satoshi KUROKAWA,
Akihiro NAKANE, Tetsuji MARUYAMA and Takahiro YASUI
Nagoya City University Graduate School of Medical Sciences, Nephro-urology, Nagoya, JAPAN
PURPOSE
To elucidate the differentiation process of spermatogonial stem cells (SSCs), we investigated the characters of SSCs and
precursor cells purified from immature rat testes exposed anti-androgen agent during fetal period. Furthermore, we also
assessed differential expressions of some stem cell markers in these cells.
MATERIAL AND METHODS
Flutamide, as anti-androgen agent, was administered to 8-week pregnant SD rats during from the 14th to20th days of
gestation via intraperitoneal injection (7.5mg/body/day). Among their male offspring, the testes at 3 to 28 days after
birth were removed. Control rats were administered only vehicle. Single-cell suspension was incubated with RPMI
medium with 5%FBS at 37°C, 5%CO2 for 24-48h. Cells of upper layer were harvested for following experiments.
Fluorescent immunohistochemistry and real-time RT-PCR were performed.
RESULTS
In fluorescent immunohistochemistry, since isolated cells were expressed Plzf, Utf1, and c-Kit proteins, these cells were
estimated SSCs. There were no significant differences of cell morphology between control and anti-androgen treatment
group. On the other hand, expression values of Plzf gene was 2.50 times, and c-Kit gene was 3.07 times in isolated cells
derived from anti-androgen treatment group compared to control.
CONCLUSIONS
Both Plzf and c-Kit are stem cell marker, and involved in self-renewal and cell proliferation of SSCs. Spermatogenic
failure was observed in the mice lacking androgen receptor (AR), and AR is expressed in the precursor cells of SSCs,
suggested that androgen has some roles in differentiation and maturation of SSCs. Suggesting that androgen is involved
in the differentiation of SSCs via regulation of these gene expressions.