Spermatogonial Stem Cells (SSCs) are unipotent adult stem cells, defined as the undifferentiated stem cells located along the basement membrane of the seminiferous tubules. This present study is aimed to carry out the culture and characterization of SSCs in pre-pubertal age group of mice. Isolation of SSCs was done by two-step enzymatic digestion method and was seeded in a six-well plate. In primary culture (P0), colony forming unit was observed on day four post incubation. Mice Spermatogonial Stem Cells (mSSCs) adherence was observed on day eight post incubation. On twelve day post incubation, the cells were subcultured for further expansion. In the subsequent passages (P1 and P2) though a heterogenous morphology was observed, most of the cells showed spindle shaped structure. The pre-pubertal mSSCs were positive for alkaline phosphatase activity at P1 and P2 showed positive expression for the transcription factor, Oct4 was indicated by intranuclear green fluorescence. Flowcytometric analysis revealed that the mSSCs in the pre-pubertal age groups showed expression for the SSEA1 marker. | In-Vitro culture and characterization of Spermatogonial stem cells SSCs in pre-pubertal mice
