The effect of macromolecular supplementation on the surface tension of TCM-199 and the utilization of growth factors by bovine oocytes and embryos in culture.
The objectives of this study were to determine the surface tension of bovine follicular fluid (BFF) and TCM-199, and the effects of the synthetic surfactant, Twin-80, or FCS, in TCM-199 on surface tension measurements and subsequent effects on bovine oocyte and embryo development. The surface tension of BFF was determined to be approximately 45.5 mN m(-1) at 25 degrees C and approximately 42.7 mN m(-1) at 39 degrees C, which was comparable to the surface tension of TCM-199 containing Twin-80 (45.7 and 43.2 mN m(-1), respectively). There was no difference in surface tension measurements of BFF from follicles 2-7 mm or 8-15 mm in diameter. Both Millipore water and phosphate buffered saline (PBS) had a surface tension measurement of 69.5 mN m(-1) at 39 degrees C. Although the presence of Twin-80 in TCM-199 resulted in a reduction in surface tension measurement as compared with unsupplemented TCM-199, there was no effect on the number of oocytes reaching metaphase II. However, the addition of epidermal growth factor (EGF) to TCM-199 containing Twin-80 did result in increased maturation rates of oocytes in vitro. There was no effect of insulin, transferrin, and selenium (ITS), or EGF on surface tension measurement of TCM-199, but significantly more zygotes cleaved and developed to morulae/blastocysts in TCM-199 containing both Twin-80 and growth factors. The addition of 20 microg EGF ml(-1) to TCM-199 containing Twin-80 was as efficacious in supporting bovine embryos in culture as was the addition of 5% or 10% fetal calf serum. This study demonstrates that surface-active components in culture media positively affect bovine oocyte maturation and embryo development in culture. Data also suggest that non-ionic surfactants, such as Twin-80 in TCM-199, may successfully replace the surface-active properties but not the embryotrophic properties of serum in embryo maturation/culture media. Although commercial TCM-199 (containing Twin-80) did not require the addition of other surface-active compounds to lower surface tension, it did benefit from the addition of growth promoting factors, which were also provided by serum.
Palasz
A T
AT
Department of Herd Medicine and Theriogenology, WCVM, University of Saskatchewan, Saskatoon, Saskatchewan, Canada. palasza@duke.usask.ca
Thundathil
J
J
Verrall
R E
RE
Mapletoft
R J
RJ
D000818:Animals; D002417:Cattle / Q000196:embryology*; D046508:Culture Techniques; D005314:Embryonic and Fetal Development / Q000187:drug effects; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005260:Female; D015571:Follicular Fluid; D006133:Growth Substances / Q000494:pharmacology*; D007328:Insulin / Q000494:pharmacology; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development*; D012643:Selenium / Q000494:pharmacology; D013499:Surface Properties; D013501:Surface-Active Agents; D014168:Transferrin / Q000494:pharmacology
D006133:Growth Substances; D007328:Insulin; D013501:Surface-Active Agents; D014168:Transferrin; D004815:Epidermal Growth Factor; D012643:Selenium