输卵管和颗粒细胞单层对牛体外受精胚胎发育的影响

Effects of bovine oviductal or granulosa cells on developmental competence of bovine IVF embryos

以屠宰场牛卵巢为试验材料,研究输卵管细胞单层(OCM)和颗粒细胞单层(GCM)对牛卵母细胞体外成熟(IVM)、体外受精(IVF)和体外培养(IVC)后胚胎发育能力的影响。(1)从卵泡抽取卵丘卵母细胞复合体(COCs),并根据卵母细胞外面卵丘细胞的层数将其分为3类:1级(≥4层);2级(2～3层);3级(0～1层)。作分别在IVM和IVC培养液中添加GCM(1×106个/mL)与不添加的对比试验。结果显示:添加GCM对1级卵母细胞的卵裂率、6～8细胞发育率和囊胚率无明显影响(P>0.05);但添加GCM的2级、3级卵母细胞,受精后的卵裂率、6～8细胞发育率和囊胚率分别高于未添加组(P<0.05)。(2)所有卵母细胞(包括COCs和裸卵)被随机分为3个组,在其IVM和IVC培养液中分别添加OCM、GCM或不添加体细胞(对照组)。结果显示:OCM和GCM组的卵裂率、6～8细胞发育率和囊胚率均高于对照组(P<0.05),而两试验组之间差异不显著。

Experiments were conducted to study the effects of oviductal cell monolayer （OCM） and granulose cell monolayel （GCM） on developmental competence of bovine oocytes derived from abattoir ovaries following in vitro maturation （IVM） , in vitro fertilization （IVF） and in vitro culture （IVC）. In experiment 1, bovine cumulus oocyte complexes （COCs） were aspirated from follicles and classified into 3 morphological categories according to the surrounding cumulus cells： Gradel （≥4 layers） , Grade2 （2 to 3 layers ） and Grade3 （0 to 1 layer）. Oocytes were co-cultured with or without granulose cells （ 1×10^6个/ml） in IVM and IVC medium. The presence of granulosea cells during maturation and culture did not affect the cleavage rate, 6 -8 cells rate and blastocyst rates （P 〉 0. 05 ） of oocytes in grade 1. However, the cleavage rate, 6～8 cells rate and blastocyst rates in grade 2 and grade 3 were higher （ P 〈 0. 05 ） when COCs were cultured in the presence of GCM than when cultured in the absence of GCM. In experiment 2, oocytes （ COCs and naked oocytes） were co-cultured with OCM, GCM or without both （control） in IVM and IVC medium. Following IVM, IVF and IVC, more oocytes in both OCM and GCM groups reached 40 cleavage, 6～ 8 cells and blastocyst than in control group （P 〈 0. 05） , although there was no significant difference （ P 〉 0. 05 ） between OCM and GCM groups.