Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

Theriogenology. 2008 May;69(8):1013-21. doi: 10.1016/j.theriogenology.2007.12.015. Epub 2008 Mar 20.

Abstract

The aim of this study was to examine the effects of co-culture with Vero cells during the in vitro maturation (IVM) and three culture media, B2+5% fetal calf serum (FCS) on Vero cells, synthetic oviduct fluid (SOF)+5% FCS, and SOF+20 gL(-1) bovine serum albumin (BSA), on the developmental competence of the embryos and their ability to survive vitrification/warming. We also tested the effect of morphological quality and the age of the embryo on its sensitivity to vitrification. The IVM system neither affects the embryo development up to Day 7 nor survival rates after vitrification. The culture of embryos in SOF+FCS and in Vero cells+B2 allowed obtaining more Day 6 and Day 7 blastocysts, and a higher % of Day 7 blastocysts vitrified than culture in SOF+BSA. Contrarily, on Day 8, more blastocysts were vitrified in SOF+BSA than in SOF+FCS. Blastocysts quality affected development after vitrification/warming, and Day 7 embryos showed higher survival rates than their Day 8 counterparts. Day 7 blastocysts produced in Vero cells or in SOF+BSA survived at higher rates than those produced in SOF+FCS at 24 and 48 h after warming. Embryo culture with BSA allows obtaining hatching rates after vitrification/warming higher than those obtained after co-culture with Vero cells in B2 and FCS. Moreover, this system provides hatching rates from Day 8 blastocysts comparable to those obtained on Day 7 in Vero cells. Further studies, including embryo transfer to recipients, are needed to clarify factors affecting the freezability of in vitro produced bovine embryos.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / physiology*
  • Cattle / physiology*
  • Chlorocebus aethiops
  • Coculture Techniques / veterinary
  • Cryopreservation / methods
  • Cryopreservation / veterinary*
  • Culture Media, Serum-Free*
  • Embryo Culture Techniques / veterinary*
  • Embryo, Mammalian
  • Embryonic Development / physiology*
  • Female
  • Fertilization in Vitro / methods
  • Fertilization in Vitro / veterinary*
  • Male
  • Serum Albumin, Bovine
  • Vero Cells

Substances

  • Culture Media, Serum-Free
  • Serum Albumin, Bovine