Study on bovine IVF zygote co-cultured with bovine fetal sertoli cells

Chinese. 研究牛胎儿睾丸支持细胞对牛早期胚胎的体外发育是否有促进作用。从屠宰场采集5~7月龄胎牛睾丸，经原代和传代培养制备牛胎儿睾丸支持细胞（sertoli cells，SCs）饲养层，对比原代和传代胎牛睾丸SCs与牛体外受精卵共培养时受精卵的发育情况；分别以4×104，2×104 mL－1两个密度接种传代胎牛睾丸SCs与牛受精卵体外共培养，并以培养液中无牛胎儿睾丸SCs饲养层为对照，研究体外牛胎儿睾丸支持细胞（sertoli cells，SCs）对牛受精卵体外发育的影响。与牛受精卵体外共培养时，接种密度为2×104 mL－1传代胎牛睾丸SCs饲养层共培养组的卵裂率（79.3%）高于原代牛胎儿睾丸SCs饲养层共培养组（69.2%），但差异不显著（P0.05）；囊胚发育率（41.3%）极显著地高于原代SCs饲养层共培养组（16.7%）（P0.01）。接种密度为4×104 mL－1传代牛胎儿睾丸SCs饲养层共培养组的卵裂率大于接种密度为2×104 mL－1传代牛胎儿睾丸SCs饲养层共培养组和对照组，但差异不显著；胚胎发育率极显著地低于接种密度为2×104 mL－1传代牛胎儿睾丸SCs饲养层共培养组和对照组。制备的传代牛胎儿睾丸SCs饲养层，能有效促进牛体外受精卵的体外发育，提高囊胚发育率；接种的传代牛胎儿睾丸SCs饲养层密度过大，将严重影响牛体外受精卵的发育。

English. In order to study the effect of bovine fetal SCs feeder layer on the development of bovine zygotes in vitro. 5―7 months bovine fetal testiculus from slaughter house was collected, bovine fetal SCs feeder layer was made by primary culture and sub-culture, then co-cultured with bovine IVF zygotes and the developing results was of zygotes compared; sub-cultured SCs was inoculated in 2×104 mL－1 and 4×104 mL－1, to carry out co-culture with bovine in vitro fertilized (IVF) zygotes, and made control group with no fetal bovine SCs feeder layer in the media, to study the effect of bovine fetal SCs feeder layer at different phase on the development of bovine zygotes in vitro. The results showed that: when IVF zygotes co-cultured with sub-cultured bovine fetal SCs (sub-culture group) in vitro, the cleavage rate of zygotes (79.3%) was higher than those co-cultured with primary cultured SCs feeder layer (primary group) (69.2 %), but there was no significant difference (P0.05); the blastocyst development rate (41.3%) was significantly higher than primary passage group (16.7%) (P0.01). Cleavage rate of zygotes co-cultured with SCs feeder layer which inoculated in 4×104 mL－1 was higher than that of in 2×104 mL－1, but there was no sifnificant differentce with that of in 2×104 mL－1, but as the lasting of the cultivation, the embryo development rate of zygotes co-cultured with SCs feeder layer which inoculated in 2×104 mL－1 was sifnificantly different, lower than that in 2×104 mL－1 and control groups. The fetal bovine SCs feeder layer made by sub-culturing cells can efficiently promote the in vitro development of bovine IVF zygotes, and increase the blastula developing rate; too high concentration of sub-culturing SCs feeder layer will effect the development of co-cultured bovine IVF zygotes obviously.