摘要
Abstract
Buffalo oocytes with compact cumulus cells were collected from slaughterhouse ovaries and cultured in TCM 199 supplemented with 5% estrus cow serum (ECS) + 0.1 μg/mL follicle stimulating hormone (FSH), prolactin (PRL 0, 0.1, 1.0, 10 μg/mL, Experiment 1), bovine follicular fluid (BFF) (0% BFF + 1.0 μg/mL PRL, 5% BFF, 5% BFF + 1.0 μg/mL PRL, Experiment 2), incubated at 38.5 ℃ in φ = 5% CO2 in humidified air. After 24 to 26 hours of maturation, IVF was done with swim-up separated frozen-thawed buffalo spermatozoa at 1×106 mL-1 in modified Tyrodes medium (TALP). At 24 to 26 hours post insemination, the oocytes were co-cultured with granulosa cell monolayer in droplets containing culture medium. The proportion of cleaved oocytes that developed to blastocyst stage within 9 days after commencing co-culture with granulosa cell monolayer was then evaluated. In experiment 1, the proportion of cleaved oocytes that developed to blastocyst stage was higher (12.8%) at 1.0 μg/mL PRL though not significantly different from the control (9.1%). In experiment 2, addition of 5% BFF to the maturation medium had significant increase in the cleavage rate of oocytes compared to the control (30.7% vs. 21.7%, P<0.05), but did not influence the proportion of cleaved oocytes that developed into blastocysts；addition of 5% BFF+1.0 μg/mL PRL to the maturation medium had a cleavage rate of 38.1%, with 14.0% of the cleaved oocytes developing to blastocysts (P<0.05). In conclusion, the results indicate that the addition of appropriate amounts of prolactin and bovine follicular fluid to the maturation medium will enhance the maturation of immature buffalo oocytes with their subsequent development to the blastocyst stage.
关键词
水牛/体外成熟/牛卵泡液/催乳素/胚泡Key words
buffalo/in vitro maturation/bovine follicular fluid/prolactin/blastocyst分类
农业科技
