Retroviral Gene Expression in Spermatogonial Stem Cells during Long-term Culture

The spermatogonial stem cell (SSCs) is unique in that it is the only cell in the adult male that can contribute genes to a subsequent generation. Permanent modification of the germ cell line may be realized if stem cells could be cultured, transfected with unique genes, and then transplanted into recipient testes. We developed a culture system that supported long-term viability of SSCs. We used a retrovirus vector (pMSCV including β-galactosidase) to stably transfect spermatogonia following long-term culture using the system developed. Expression of the reporter gene β-galactosidase controlled by the retroviral vector was stable in long-term cultured SSCs. We confirmed the retroviral-mediated β-galactsidase gene could be expressed in germ cells in recipient mice following SSCs transplantation. (