Effect of culture media on embryo development from prepubertal goat IVM-IVF oocytes

Theriogenology. 1999 Oct 1;52(5):847-61. doi: 10.1016/S0093-691X(99)00177-6.

Abstract

Experiments were carried out to develop and improve in vitro culture systems for IVM-IVF prepubertal goat oocytes. Cumulus oocyte complexes (COC) were obtained by slicing ovaries from slaughtered prepubertal goats. Oocytes were matured in TCM-199 supplemented with 20% estrous goat serum (EGS) + 10 micrograms/mL FSH + 10 micrograms/mL LH + 1 microgram/mL estradiol 17 beta for 27 h at 38.5 degrees C in 5% CO2 in air. Matured oocytes were placed in drops of TALP- fert medium supplemented with hypotaurine (1 microgram/mL) and inseminated with freshly ejaculated spermatozoa following capacitation as described by Younis et al. (69) but with 100 micrograms/mL heparin. At 24 h post insemination the ova were transferred to various in vitro culture media, and early embryo development was evaluated until Day 8 post insemination. Specifically, in the studies described here, we have compared the effects of (Experiment 1) co-culture systems using oviductal ephitelial cells (OEC) and cumulus cells (CC), both caprine and bovine; (Experiment 2) the presence of serum and/or OEC; (Experiment 3) 4 culture media (TCM199, Ham's F10, CZB abd SOF) for co-culture with OEC; and (Experiment 4) conditioned medium with OEC. In Experiment 1, the percentage of morulae plus blastocysts was higher in culture with OEC, both caprine and bovine (15.1 and 14.8%, respectively) than with CC (4.1 and 6.7%, respectively). In Experiment 2, the OEC with EGS did not improve the percentage of morulae and blastocysts obtained with OEC alone (14.3 and 23.1% respectively). In Experiment 3, this percentage was higher using OEC with TCM-199 compared to CZB medium (21.3 and 12.3%, respectively) and in Experiment 4, the results were 3.7, 11.2 and 21.3% for TCM-199 without cells, Conditioned Medium and co-culture with OEC, respectively.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blastocyst / cytology
  • Blastocyst / physiology*
  • Cattle
  • Cells, Cultured
  • Coculture Techniques
  • Culture Media
  • Embryonic and Fetal Development / physiology*
  • Epithelial Cells / cytology
  • Female
  • Fertilization in Vitro / methods
  • Fertilization in Vitro / veterinary
  • Goats
  • Heparin / pharmacology
  • Male
  • Morula / cytology
  • Morula / physiology*
  • Oocyte Donation / veterinary
  • Oocytes / cytology*
  • Oocytes / physiology
  • Ovary / cytology
  • Sexual Maturation
  • Spermatozoa / drug effects
  • Spermatozoa / physiology
  • Taurine / analogs & derivatives

Substances

  • Culture Media
  • Taurine
  • hypotaurine
  • Heparin