Experimental Investigation of Ki67, POU5F1, and ZBTB16 Expression in the Pig and Mouse Testicular Cells using Immunocytochemistry and RT-PCR

Introduction: Spermatogonial Stem Cells (SSC) are the originators and beginning points of the spermatogenesis process. Moreover, they are considered the only stem cells in the body that could transfer genetic information to the next generation through gametogenesis. This study aimed to investigate the potency and power of SSC under in vitro and in vivo conditions.   Materials & Methods:  Enzymatic digestion technique was utilized to extract the spermatogonial cells of the pig and mousechr('39')s testis. They were then cultured in an environment containing FGF, EGF, GDNF, and a feeder layer of STO.  For immunocytochemistry and RT-PCR analysis, Ki67, POU5F1, and ZBTB16 markers were used to evaluate the resulted colonies. Ethics code: Ir.ausmt.rec.1398.03.07     Findings: The nature of the SSC resulted after separation and culture was proved through measures, such as cluster growth of the colonies in the culture medium, Ki67 marker expression in the immunocytochemistry review which showed the duplication ability, and the morphological criteria observed by an electron microscope. Moreover, the comparative expression of POU5F1 and ZBTB16 markers in the embryonic stem cells, SSC, and Sertoli cells within the seminiferous tubules of the mouse was analyzed by RT-PCR.   Discussions & Conclusions: This experimental study investigated the expression of Ki67, POU5F1, and ZBTB16 in the seminiferous tubules and special cytological features of SCC. The findings are beneficial for future advanced studies in reproductive biology fields.