Short-term in-vitro culture of goat enriched spermatogonial stem cells using different serum concentrations

J Assist Reprod Genet. 2012 Jan;29(1):39-46. doi: 10.1007/s10815-011-9687-5. Epub 2011 Dec 11.

Abstract

Purpose: To investigate the effect of serum supplementing on short-term culture, fate determination and gene expression of goat spermatogonial stem cells (SSCs).

Methods: Crude testicular cells were plated over Datura-Stramonium Agglutinin (DSA) for 1 h, and non-adhering cells were cultured in the presence of different serum concentrations (1, 5, 10, and 15%) for 7 days in a highly enriched medium initially developed in mice. Colonies developed in each group were used for the assessment of morphology, immunocytochemistry, and gene expression.

Results: Brief incubation of testicular cells with DSA resulted in a significant increase in the number of cells that expressed the germ cell marker (VASA). The expression of THY1, a specific marker of undifferentiated spermatogonia, was significantly higher in colonies developed in the presence of 1% rather than 5, 10 and 15% serum.

Conclusion: Goat SSCs could proliferate and maintain in SSC culture media for 1 week at serum concentrations as low as 1%, while higher concentrations had detrimental effects on SSC culture/expansion.

MeSH terms

  • Agglutinins / chemistry
  • Animals
  • Cell Culture Techniques / methods*
  • Cell Differentiation / genetics
  • Culture Media / chemistry*
  • Datura stramonium / chemistry
  • Gene Expression
  • Goats / genetics*
  • Male
  • Mice
  • Serum / chemistry
  • Spermatogonia / cytology*
  • Stem Cells / cytology*
  • Thy-1 Antigens / metabolism

Substances

  • Agglutinins
  • Culture Media
  • Thy-1 Antigens