牛体外受精胚胎衍生干细胞能力影响因素的研究

以牛卵巢卵母细胞体外受精获取囊胚期胚胎，比较体外受精牛胚胎不同获取内细胞团的方法和不同培养液对体外培养胚胎干细胞能力的影响。结果表明，囊胚期胚胎不除去透明带而直接培养产生胚胎干细胞，初次克隆率为55％；胰酶法去透明带分离的内细胞团（ICM）培养产生胚胎干细胞，初次克隆率为90％。免疫外科法去透明带分离的ICM在4种不同的培养液中，初次克隆率分别为16．4％、11．5％、21．8％、18．2％，但是其分离的ICM经传代后形成的衍生细胞不易发生分化，经过4～6代的传代，仍然保持其完整的形态，说明免疫外科法是一种理想的牛ICM分离法，培养液为D20＋LIF（40ng／mL）可用于牛胚胎干细胞培养。

Effect of Several Factors on Developmental Potential of Bovine Embryonic Stem Cells from IVF Embryo

The objectives of this study were to derive bovine embryonic stem cells from the inner cell mass of IVF blastocysts and to learn the effect of several factors on developmental potential of bovine embryonic stem cells from IVF embryo. The results showed that., the primary colony rates of bovine embryonic stem cell from normal IVF blastocysts and trypsin-isolated ICMs were 55% and 90%, respectively. Embryonic stem cell from immunosurgical-isolated ICM and cultured in four media, the primary colony rates were 16.4% ,11.5% ,21.8% and 18.2% ,respectively. The embryonic stem cell from immunosurgical-isolated ICM have been kept undifferentiated, from which the individual cells morpha and growth were better than that of stem-cells from trypsin-isolated ICMs. Finally the results demonstrated that the immunosurgical-isolated ICM was a suitable isolated method.