Expression of Spermatogonial and Pluripotency Markers in Spermatogonial Stem Cells after Treatment with Different Culture Factors

As condition and component of culture determine fate map of spermatogonial stem cells (SSCs), the aim of this study was to evaluate of growth factors GDNF, LIF and RA on proliferation and differentiation of SSC. SSCs were cultured in two groups: The first group GDNF and LIF and the second group RA. The number of clumps and colony formation was monitored during 1 month in culture. To identification of the colony, stained with PLZF using immunostaining. Pluripotency gene Oct 4 and neural markers MAP2, NeuroD and Nestin were analyzed by RT-PCR. In the presence of GDNF and LIF, cells proliferated rapidly and many compact clumps were appeared whereas after exposure to RA cells formed small clumps. The results of immunocytochemistry shows PLZF was detected in the group GDNF & LIF. RT-PCR showed high level expression Oct 4 in the group GDNF and LIF whereas neural markers MAP2, NeuroD and Nestin were expressed in the group RA. GDNF and LIF are essential for self-renewal and colony formation of SSCs that confirm the stem cells activity of these cells but RA inhibits stem cell activity of SSCs and induces neural differentiation of these.