Bernardi M L, Fléchon J E, Delouis C
Génétique Moléculaire et Cellulaire, INRA-ENVA, Maisons-Alfort, France.
J Reprod Fertil. 1996 Mar;106(2):161-7. doi: 10.1530/jrf.0.1060161.
The development and quality of ovine zygotes matured and fertilized in vitro were compared after coculture with oviductal cells (CZB-199 system) and culture in synthetic oviduct fluid medium without cells (SOF system). The effect of two oxygen concentrations (5% and 20%) on the development of ovine zygotes in SOF medium was also studied. More ovine zygotes reached the blastocyst stage when culture in SOF medium was performed in 5% O2 rather than 20% O2. A greater number of blastocysts was obtained after culture in the SOF system than coculture in the CZB-199 system. Proportions of grade I (excellent), II (good), III (fair) and IV (poor) blastocysts did not differ significantly between the SOF and CZB-199 systems. Histological examination of hatched blastocysts revealed a superiority of the SOF system for the following: a greater number of total and trophoblastic cells in grade I and II blastocysts; more endodermic cells in grade I blastocysts, higher mitotic index in the inner cell mass of grade II blastocysts and in total and trophoblastic cells of grade I, II and III blastocysts; more grade III blastocysts with mitosis in the inner cell mass; and a lower pyknotic index in the inner cell mass of grade I, II and III blastocysts. Culture in the SOF system improved the rate and quality of blastocysts in comparison with the CZB-199 system. Furthermore, culture in SOF medium with 5% O2 provided more blastocysts than culture in the presence of 20% O2.
将体外成熟和受精的绵羊受精卵分别与输卵管细胞共培养(CZB - 199系统)以及在无细胞的合成输卵管液培养基中培养(SOF系统),比较二者的发育情况及质量。同时研究了两种氧气浓度(5%和20%)对SOF培养基中绵羊受精卵发育的影响。当在5%氧气环境而非20%氧气环境中进行SOF培养基培养时,更多的绵羊受精卵能发育到囊胚阶段。与CZB - 199系统共培养相比,在SOF系统中培养后获得的囊胚数量更多。在SOF系统和CZB - 199系统中,I级(优秀)、II级(良好)、III级(中等)和IV级(差)囊胚的比例没有显著差异。对孵化囊胚的组织学检查显示,SOF系统在以下方面具有优势:I级和II级囊胚中的总细胞数和滋养层细胞数更多;I级囊胚中的内胚层细胞更多;II级囊胚内细胞团以及I级、II级和III级囊胚的总细胞和滋养层细胞中的有丝分裂指数更高;内细胞团中有有丝分裂的III级囊胚更多;I级、II级和III级囊胚内细胞团中的固缩指数更低。与CZB - 199系统相比,在SOF系统中培养提高了囊胚的发育率和质量。此外,与在20%氧气环境中培养相比,在含5%氧气的SOF培养基中培养能获得更多囊胚。