Advanced isolation, expansion and characterization research study on pig testicular cells during differentiation and proliferation

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 pig and consequently has an impact on the analysis of pig fertility. Germ cells were extracted from pig 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 pig testicular cells. Electron microscopy was also utilized to analyze the morphological features of the extracted pig germ cells. 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. The heterogeneity of the in vitro cultured cells was detected via morphological analysis by the electron microscope. In this experimental study, we tried to reveal exclusive information which obviously could be helpful for future success in the achievement of proper therapies against infertility and sterility as an important global issue.