Evaluation Of Porcine Somatic And Testicular Germ Cells During Differentiation And Proliferation

Abstract

Background: Spermatogenesis is the complex process of sperm production to transmit paternal genetic information to the subsequent generation. This process is determined by the collaboration of several germ and somatic cells, most importantly spermatogonia stem cells and Sertoli cells. To characterize germ and somatic cells in the tubule seminiferous contort in porcine and consequently has an impact on the analysis of porcine fertility.

Materials and methods: Germ cells were extracted from porcine testis by enzymatic digestion before being expanded on Sandos inbred mice (SIM) embryo-derived thioguanine and ouabain-resistant fibroblasts (STO) feeder layer supplemented with FGF, EGF, and GDNF. Immunohistochemistry (IHC) and immunocytochemistry (ICC) analysis for Sox9, Vimentin, and PLZF markers were performed to examine the generated colonies of porcine testicular cells. Electron microscopy was also utilized to analyze the morphological features of the extracted porcine germ cells.

Results: IHC analysis revealed that Sox9 and Vimentin were expressed in the basal compartment of the seminiferous tubules. Moreover, ICC results showed that the cells have low expression of PLZF while expressing vimentin. Heterogeneity of the in vitro cultured cells was detected via morphological analysis by the electron microscope.

Conclusion: In this experimental study, we tried to reveal the exclusive information which obviously could be helpful for future success in achievement to proper therapies against infertility and sterility as an important global issue.