牛体外受精胚胎无血清培养的研究  

Study on in vitro Cultural System of bIVF Embryos in Serumfree Medium

作　　者：高建明[1] 朱裕鼎[2] 刘云海[2] 敖红[2] 杨文胜[2] 

机构地区：[1]北京农学院 [2]中国农业科学院畜牧研究所

出　　处：《中国农业科学》1998年第3期79-85,共7页Scientia Agricultura Sinica

基　　金：北京市青年科技骨干培养基金

摘　　要：以ＴＣＭ１９９为基础液，对常规ＩＶＭ／ＩＶＦ牛早期胚胎进行了无血清（１ｍｇ／ｍｌ，ＰＶＡ，聚乙烯醇）和有血清（２０％ＦＣＳ）培养的对比实验。通过在无血清培养液中加入９种添加物，初步筛选出了在５％ＣＯ２、９５％空气、３９℃、饱和湿度培养环境下的牛体外受精胚胎无血清培养体系为：ＴＣＭ１９９＋ＥＤＴＡ０．１ｍｍｏｌ／Ｌ＋Ｉ５μｇ／ｍｌ＋Ｔ５μｇ／ｍｌ＋Ｓ５ｎｇ／ｍｌ＋ＥＧＦ１０ｎｇ／ｍｌ＋牛磺酸７ｍｍｏｌ／Ｌ＋亚牛磺酸５ｍｍｏｌ／Ｌ＋ＰＶＡ１ｍｇ／ｍｌ＋青霉素１００ｉｕ／ｍｌ＋链霉素１００ｍｇ／ｍｌ。无血清培养与有血清培养囊胚率（７．１％对１４．３％）、扩展囊胚率（４．３％对７．１％），均无显著差异（Ｐ＞０．０５）。实验结果还表明，外源激素（雌激素和孕酮）对胚胎发育无促进作用；有血清组与无血清组胚胎的各发育阶段（桑椹胚、囊胚、扩展囊胚）细胞核数量无明显差异（Ｐ＞０．０５）。Early bovine embryos routinely obtained from IVM/IVF oocytes, were cultured in two culture media based on tissue culture medium TCM199: serumfree one (TCM199+1mg/ml PVA) and seruminvolved one (TCM199 with 20% FCS). In comparison with TCM199+20%FCS, serumfree medium was modified step by step for culture of bovine IVM/IVF embryos to the blastooyst stage, a serumfree complex culture medium was harvested, whose constituent included TCM199+0.1mmol/L EDTA+5μg/ml Insulin+5μg/ml Transferrin+5ng/ml Sodium Selenite+10ng/ml Epidermal Growth Factor+7mmol/L Taurine+5mmol/L Hypotaurine+1mg/ml PVA+100iu/ml Penicillin+100mg/ml Streptomycin, thought to be suitable for development of early bovine embryos in an atmosphere of 5%CO2, 95%air with saturated humidity at 39℃. There was no difference between serumfree complex medium and TCM199+20%FCS in supporting embryos to blastocyst and expanded blastocyst stages of development (7.1%vs14.3%; 4.3%vs7.1%). Additional experimental data also indicated that addition of exogenous hormones (estrogen and progesterone) shed no light on improvement of development of early bovine embryos. Furthermore, the mean number of nuclei per embryo at the stage of morula, or blastocyst or expanded blastocyst, was of no significant difference for developing in either serumfree or serumimplicated medium (P>0.05).

关 键 词：牛 体外受精 胚胎 无血清 体外培养 PVA 

分 类 号：S823.34[农业科学—畜牧学]