Aim: 

Several studies have recently shown that in vitro manipulation can transform mouse and human spermatogonial stem cells (SSCs) into pluripotent stem cells, opening new opportunities in the field of regenerative medicine. Here we derived and characterized pluripotent stem cells from mouse adult testes.

Methods: 

Cells enzymatically isolated from seminiferous tubules were cultured in the presence of glial-derived neurotrophic factor (GDNF) and leukaemia inhibitory factors (LIF), important for SSCs survival and proliferation. After 4 days cells were plated on a layer of embryonic fibroblasts (STO) in ESCs culture medium; after 4-5 weeks, colonies with a typical ESC-like morphology appeared.

Results: 

RT-PCR data showed that SSC-derived colonies expressed the transcription factors Oct4 and Nanog and the alkaline phosphatase features typical of ESCs, supporting cell pluripotency. Flow cytometry analysis carried out on both ESCs and SSC-derived cells showed a similar pattern of the stem cell markers SSEA1, Oct4, Nanog, CD90, Sca1, c-kit, CD49f and CD34. SSC-derived ES-like cells when differentiated through the formation of embryoid bodies (EBs), which recapitulate the early developmental stages of the embryo, developed foci of spontaneously contracting cells, demonstrating differentiation toward a cardiac phenotype.

Conclusion: 

RT-PCR and immunofluorescence analysis confirmed the ability of these cells to form derivatives of the three germ layers.