Germ cell differentiation of bone marrow mesenchymal stem cells

Andrologia. 2019 May;51(4):e13229. doi: 10.1111/and.13229. Epub 2019 Feb 11.

Abstract

Bone marrow mesenchymal stem cells (BM-MSCs) were first cultured under induction of retinoic acid (RA), Sertoli cells conditioned medium and RA + con (conditioned medium) as treatment groups. The presence of Sertoli cells was confirmed by immunocytochemistry of follicle-stimulating hormone receptor in Sertoli cells and flow cytometry by anti-Gata4 antibody. Cell viability and morphology of nucleus and cytoplasm of BM-MSCs were evaluated by MTT test and DAPI staining respectively. The expression of Oct4, Plzf, Scp3, Caspases 8, 9 and 3 genes was evaluated by RT-PCR. For increasing the accuracy of experiment, the expression of Vasa and SCP3 genes was investigated quantitatively by real-time PCR after 0, 5, 10, 15 days of culture. The results showed that the number of apoptotic cells increased in RA group. The expression of apoptosis genes (Caspases 3, 8 and 9) was also observed in this group all days of culture. Measurement of Vasa and Scp3 genes by RT-PCR confirmed the positive effects of retinoic acid on increasing of genes expression. So, in this study, a group with maximum expression of differentiation genes and minimum expression of apoptotic genes was RA + conditioned medium group. DNA fragmentation was not observed in all groups.

Keywords: BM-MSCs; apoptosis; differentiation; spermatogenesis.

MeSH terms

  • Animals
  • Bone Marrow Cells / physiology*
  • Cell Culture Techniques / methods
  • Cell Differentiation*
  • Cell Survival
  • Cells, Cultured
  • Culture Media, Conditioned
  • Flow Cytometry
  • Germ Cells / physiology*
  • Infertility, Male / therapy
  • Male
  • Mesenchymal Stem Cells / physiology*
  • Mice
  • Sertoli Cells
  • Tretinoin / metabolism

Substances

  • Culture Media, Conditioned
  • Tretinoin