Influence of culture medium and age of bovine blastocysts in established colonies of embryonic stem cells

J Stem Cells. 2014;9(4):225-34.

Abstract

The objective of this study was to evaluate the potential of bovine IVF blastocysts at different stages of embryonic development in establishing ESC-like. Furthermore, blastocysts cultured in medium containing (10%) and (2.5%) fetal calf serum (FCS) were compared to determine if the serum concentration during in vitro culture alters the blastocyst's potential to establish ESC-like culture. It was observed that only ICM's from expanded blastocysts adhered to the monolayer (n=160) independent of the concentration of serum used during IVF culture. There were no observable differences in potential to establish ESC-like in embryos cultured with 2,5% or 10% FCS . The bFGF didn´t seems to be required for maintenance of bovine ESC-like regardless of culture conditions. Blastocysts and colonies in primary culture and after the first passage were positive for Oct4, Nanog, SSEA-3 and TRA-1-81. Expanded blastocysts gave rise to ESC-like colonies, and the addition of LIF was sufficient to maintain cells undifferentiated in culture.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / cytology*
  • Blastocyst / metabolism
  • Cattle
  • Cell Culture Techniques / methods*
  • Culture Media / chemistry*
  • Embryonic Stem Cells / cytology*
  • Fertilization in Vitro
  • Fibroblast Growth Factors / metabolism

Substances

  • Culture Media
  • Fibroblast Growth Factors