The PI, Dr. Dirk de Rooij, requests three years of support to investigate factors influencing spermatogonial stem cell growth and differentiation in culture. Spermatogonial stem cells will be purified from testes of vitamin A-deficient (VAD) mice.
The first aim i s to develop a culture system for maintaining long-term viability of these highly purified and undifferentiated spermatogonia.
The second aim i s to develop culture conditions that will promote proliferation of spermatogonia.
The third aim i s to establish immortalized cell lines of undifferentiated cells, using transfection with immortalizing genes if necessary.
The fourth aim i s to find the factor(s) that will induce differentiation of the cultured spermatogonia.
The final aim i s to test the developmental competence of these cells by studying the effects of various intervals and conditions of culture on the ability of the spermatogonial stem cells (and stem cell lines) to repopulate germ cell-deficient testes after transplantation.
| Creemers, L B; Meng, X; den Ouden, K et al. (2002) Transplantation of germ cells from glial cell line-derived neurotrophic factor-overexpressing mice to host testes depleted of endogenous spermatogenesis by fractionated irradiation. Biol Reprod 66:1579-84 |
| de Rooij, D G; Russell, L D (2000) All you wanted to know about spermatogonia but were afraid to ask. J Androl 21:776-98 |
