Isolation and In vitro Culture of Pig Spermatogonial Stem Cell

Han, Su Young; Gupta, Mukesh Kumar; Uhm, Sang Jun; Lee, Hoon Taek

doi:2009.22.2.187

Article

Animal Reproduction and Physiology

Asian-Australasian Journal of Animal Sciences 2009;22(2): 187-193.
https://doi.org/10.5713/ajas.2009.80324 Published online January 6, 2009.

Isolation and In vitro Culture of Pig Spermatogonial Stem Cell

Su Young Han, Mukesh Kumar Gupta, Sang Jun Uhm, Hoon Taek Lee^*

Correspondence: Hoon Taek Lee,

Abstract

The present study identified the favorable conditions for isolation, enrichment and in vitro culture of highly purified, undifferentiated pig spermatogonial stem cell (SSC) lines that proliferate for long periods of time in culture. The colonies displayed morphology similar to miceSSC and were positive for markers of SSC (PGP9.5), proliferating germ cell (PigVASA), pre-meiotic germ cell (DAZL) and pluripotency (OCT4, SSEA-1, NANOG, and SOX2) based on immuno-cytochemistry and RT-PCR. The purity of these colonies was confirmed by negative expression of markers for sertoli cell (GATA4 and SOX9), peritubular myoid cell (-SMA), differentiating spermatogonial and germ cells (c-KIT). The colonies could be maintained with undifferentiated morphology for more than two months and passaged more than 8 times with doubling time between 6-7 days. Taken together, we conclude that pigSSC could be successfully isolated and cultured in vitro and they possess characteristics similar to miceSSC.

Keywords: Spermatogonial Stem Cell; SSC; Pluripotency; Long Term Culture; Pig