水牛活体采卵和无透明带胚胎培养体系的优化

doi:10.16431/j.cnki.1671-7236.2020.12.024

Abstract

Abstract: The purpose of this study was to explore the problems of low development efficiency of small amounts in vitro fertilization (IVF) embryos and limited development potential of zona pellucida-free embryos when cultured in vitro of mammals,to establish an efficient and stable in vitro production system for improving the developmental potential of living ovum-pick-up (OPU) and handmade clone (HMC) embryos in buffalos.The study first compared the effect of the number of fertilized eggs (1,3,5,10 and 20) co-cultured within a single microdroplet on the embryonic development effect.Then,OPU-IVF embryos were cultured using the well-of-the-well (WOW) system and the auxiliary co-culture system (adding agarosaccharide fragments of embedded in in vitro fertilization (IVF) fertilized egg in the cultivation of microdroplets).Furthermore,the embryos without zona pellucida were cloned and reconstructed by using the WOW system and compared with the traditional microdroplet system in vitro.The results showed that the blastocyst development rates of the 10 and 20 embryos groups were significantly higher than that of the 1,3 and 5 embryos groups.Compared with the microdroplet system,the assisted co-culture system and the WOW system significantly improved the blastocyst rate of OPU-IVF embryos(P<0.01).Moreover,the WOW culture system significantly promoted the cleavage rate and blastocyst rate of HMC reconstructed embryos(P<0.01).To sum up,embryo mass culture contributed to embryo development,and under the premise of ensuring a clear pedigree,the agar-sugar embedding assisted embryo co-culture system and the WOW system improved the in vitro development efficiency of OPU-IVF embryos,the WOW system would also be applied to the high-efficient culture of zona pellucida free embryos.

Key words: buffalos; ovum-pick-up(OPU); handmade clone (HMC); agarose-embedded helper; well-of-the-well system

CLC Number:

S813.7

YANG Chunyan, ZHENG Haiying, LI Lingyu, HUANG Jiaxiang, HUANG Chunli, SHANG Jianghua. Optimization of in vitro Culture System for Embryos Derived from Ovum-pick-up and Zona-free Embryos in Buffalos[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(12): 4008-4015.

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Optimization of in vitro Culture System for Embryos Derived from Ovum-pick-up and Zona-free Embryos in Buffalos

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