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

Abstract

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.

Keywords:

• Pig
• reproduction
• seminiferous tubule
• sperm

Authors’ contributions

Mehdi Mehdinezhad Roshan: Carried out and design the experiment, assembly of data and data analysis, wrote the manuscript and data analysis; Hossein Azizi.; Carried out and provided critical feedback.

Ethical approval

The study was approved by the Ethics Committee of Amol University of Special Modern Technologies (Ir.ausmt.rec.1398.03.07).

Disclosure statement

No potential conflict of interest was reported by the author(s).

Data availability statement

The data sets analyzed for the current study are available from the corresponding author on reasonable request.

Additional information

Funding

No funding was received.