title: 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.
abstract: 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.
authors.affiliation: Department of Herd Medicine and Theriogenology, WCVM, University of Saskatchewan, Saskatoon, Saskatchewan, Canada. palasza@duke.usask.ca
authors.forename: A T
authors.initials: AT
authors.lastname: Palasz
authors.forename: J
authors.initials: J
authors.lastname: Thundathil
authors.forename: R E
authors.initials: RE
authors.lastname: Verrall
authors.forename: R J
authors.initials: RJ
authors.lastname: Mapletoft
mesh_terms: 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
chemical_list: D006133:Growth Substances; D007328:Insulin; D013501:Surface-Active Agents; D014168:Transferrin; D004815:Epidermal Growth Factor; D012643:Selenium