In vitro culture alters cell lineage composition and cellular metabolism of bovine blastocyst†

Biol Reprod. 2024 Jul 12;111(1):11-27. doi: 10.1093/biolre/ioae031.


Profiling bovine blastocyst transcriptome at the single-cell level has enabled us to reveal the first cell lineage segregation, during which the inner cell mass (ICM), trophectoderm (TE), and an undefined population of transitional cells were identified. By comparing the transcriptome of blastocysts derived in vivo (IVV), in vitro from a conventional culture medium (IVC), and in vitro from an optimized reduced nutrient culture medium (IVR), we found a delay of the cell fate commitment to ICM in the IVC and IVR embryos. Developmental potential differences between IVV, IVC, and IVR embryos were mainly contributed by ICM and transitional cells. Pathway analysis of these non-TE cells between groups revealed highly active metabolic and biosynthetic processes, reduced cellular signaling, and reduced transmembrane transport activities in IVC embryos that may lead to reduced developmental potential. IVR embryos had lower activities in metabolic and biosynthetic processes but increased cellular signaling and transmembrane transport, suggesting these cellular mechanisms may contribute to improved blastocyst development compared to IVC embryos. However, the IVR embryos had compromised development compared to IVV embryos with notably over-active transmembrane transport activities that impaired ion homeostasis.

Keywords: bovine blastocyst; in vitro culture; lineage specification; reduced nutrient medium; single-cell RNA-seq.

MeSH terms

  • Animals
  • Blastocyst* / cytology
  • Blastocyst* / metabolism
  • Cattle
  • Cell Lineage*
  • Culture Media
  • Embryo Culture Techniques* / veterinary
  • Embryonic Development / physiology
  • Female
  • Transcriptome


  • Culture Media