Effects of different Sertoli cell types on the maintenance of adult spermatogonial stem cells in vitro

In Vitro Cell Dev Biol Anim. 2017 Sep;53(8):752-758. doi: 10.1007/s11626-017-0172-z. Epub 2017 Jul 11.

Abstract

Spermatongonial stem cells (SSCs) are unique testis cells that are able to proliferate, differentiate, and transmit genetic information to the next generation. However, the effect of different Sertoli cell types on the expression of specific SSC genes is not yet well understood. In this study, we compare the in vitro effect of adult Sertoli cells, embryonic Sertoli cells, and TM4 (a Sertoli cell line) as feeder layers on the expression of SSC genes. SSCs were isolated from the testis of adult male mice and purified by differential plating. Following enrichment, SSCs were cultivated for 1 and 2 wk in the presence of various feeders. The expression of SSC-specific genes (Mvh, ZBTB, and c-kit) was evaluated by real-time polymerase chain reaction. Our results revealed that expression of the specific SSC genes was significantly higher in the embryonic Sertoli cells after 1 and 2 wk compared to the adult Sertoli cells and the TM4 group. Our finding suggest that co-culturing of SSCs with embryonic Sertoli cells is helpful for in vitro cultivation of SSCs and might improve the self-renewal of these stem cells.

Keywords: Culture; Feeder; Propagation; Sertoli cell; Spermatogonial stem cell.

MeSH terms

  • Adult Germline Stem Cells / cytology*
  • Adult Stem Cells / cytology*
  • Animals
  • Cell Lineage / genetics
  • Cell Proliferation / genetics*
  • Cell Self Renewal / genetics
  • DEAD-box RNA Helicases / genetics
  • Gene Expression Regulation, Developmental
  • Male
  • Mice
  • Proto-Oncogene Proteins c-kit / genetics
  • Repressor Proteins / genetics
  • Sertoli Cells / cytology*
  • Spermatogonia / cytology
  • Spermatogonia / growth & development
  • Testis / cytology
  • Testis / growth & development

Substances

  • Repressor Proteins
  • ZBTB1 protein, mouse
  • Proto-Oncogene Proteins c-kit
  • Ddx4 protein, mouse
  • DEAD-box RNA Helicases