Factors affecting the in-vitro development to blastocysts of bovine oocytes matured and fertilized in vitro
Abstract
Summary. The effects of media (TCM199 vs. synthetic oviduct fluid, SOF), sera (foetal calf serum, FCS vs. human serum, HS), gas atmosphere (5% CO2 in air vs. 5% CO2, 5% O2 and 90% N2) and coculture with bovine oviduct epithelial cells (cells vs. no cells) on the in-vitro development of in-vitro matured and fertilized bovine oocytes were examined. Immature oocytes surrounded with compacted cumulus cells were cultured for 24 h in TCM199 supplemented with 10% FCS, 10 μg follicle-stimulating hormone (FSH)/ml and 10 μg luteinizing hormone (LH)/ml, 1 μg oestradiol/ml, and 1 × 106 granulosa cells at 39°C under 5% CO2 in air. In-vitro fertilization was performed with frozen–thawed heparin-treated (100 μg/ml, 15 min) spermatozoa from 2 bulls. Oocytes were incubated with 2·5 × 106 spermatozoa/ml for 24 h and then cultured in one of 16 treatments for 7 days. Cleavage (2–8-cell) and development to blastocysts were recorded on Days 2 and 7, respectively, after the start of culture. SOF was superior to TCM199 for cleavage (P < 0·01), development to blastocysts (P < 0·001) and for proportion of cultured ova resulting in blastocysts with at least 60 or at least 100 nuclei (P < 0·001). FCS was superior to HS for development to blastocysts (P < 0·001) and 5% oxygen was superior to air for the proportion of ova reaching at least 60 cells (P < 0·01). For cleavage and development to blastocysts, there was an interaction between serum and cells (P < 0·01). In the presence of cells, ova preferred…
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