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水牛(Bubalus bubalis)卵母细胞的体外成熟与受精以及随后胚胎的发育:卵母细胞质量和血清类型的影响

In vitro maturation and fertilization, and subsequent development of buffalo (Bubalus bubalis) embryos: effects of oocyte quality and type of serum.

作者信息

Chauhan M S, Singla S K, Palta P, Manik R S, Madan M L

机构信息

Embryo Biotechnology Centre, National Dairy Research Institute, Karnal, India.

出版信息

Reprod Fertil Dev. 1998;10(2):173-7. doi: 10.1071/r97080.

Abstract

In Experiment 1, to determine the developmental potential of buffalo oocytes of different qualities, compact cumulus-oocyte complexes (COCs) with an unexpanded cumulus mass, and with homogeneous ooplasm were classified as Grade 1 (with > or =5 layers of cumulus cells) and Grade 2 less than 4 layers of cumulus cells). Grade-3 oocytes were either without cumulus cells or with expanded cumulus mass, and with irregular ooplasm. The oocytes were matured for 24 h at 38.5 degrees C, 5% CO2 in air in maturation medium (10% fetal bovine serum (FBS) in TCM-199 supplemented with 5 microg mL(-1) follicle stimulating hormone-P). The nuclear maturation and cleavage rates, and the proportion of cleaved embryos which developed to morula and blastocyst stage were in the order Grade 1>Grade 2>Grade 3 (P < 0.05). For Experiment 2, the maturation medium consisted of TCM-199 supplemented with one of the following sera at 10% concentration: (1) buffalo oestrus serum (BOS), (2) superovulated buffalo serum (SBS), (3) fetal bovine serum (FBS) and (4) steer serum (SS). After in vitro fertilization (IVF), the oocytes were co-cultured with buffalo oviductal epithelial cells in TCM-199 containing the respective sera at 10% concentration for the subsequent 9 days. The extent of cumulus expansion and nuclear maturation were not different among different groups. The cleavage rates were lower (P < 0.05) with FBS than with BOS, SBS and SS. The proportion of cleaved embryos which developed to blastocyst stage was higher (P < 0.05) with SBS than with BOS, FBS and SS.

摘要

在实验1中,为了确定不同质量水牛卵母细胞的发育潜力,将卵丘致密、卵丘团未扩张且卵质均匀的卵丘-卵母细胞复合体(COCs)分为1级(卵丘细胞层数≥5层)和2级(卵丘细胞层数少于4层)。3级卵母细胞要么没有卵丘细胞,要么卵丘团扩张,且卵质不规则。将卵母细胞在含有5μg/mL促卵泡激素-P的TCM-199(添加10%胎牛血清(FBS))的成熟培养基中,于38.5℃、5%二氧化碳的空气中成熟培养24小时。核成熟率、卵裂率以及发育至桑椹胚和囊胚阶段的卵裂胚胎比例依次为1级>2级>3级(P<0.05)。在实验2中,成熟培养基由添加了以下10%浓度血清之一的TCM-199组成:(1)水牛发情期血清(BOS)、(2)超排水牛血清(SBS)、(3)胎牛血清(FBS)和(4)公牛血清(SS)。体外受精(IVF)后,将卵母细胞与水牛输卵管上皮细胞在含有各自10%浓度血清的TCM-199中共同培养9天。不同组之间卵丘扩张程度和核成熟情况没有差异。与BOS、SBS和SS相比,FBS组的卵裂率较低(P<0.05)。与BOS、FBS和SS相比,SBS组发育至囊胚阶段的卵裂胚胎比例较高(P<0.05)。

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