【摘要】 目的 :探讨布法罗大鼠肝细胞条件培养基 (Buffaloratlivercellconditionedmedium ,BRL)在ES细胞培养中的应用方法。方法 :ES细胞复苏后分别培养在BRL条件培养基、小鼠胚胎成纤维细胞饲养层 (mouseembryonicfibroblast,MEF)及合并应用BRL条件培养基和MEF饲养层的环境中 ,通过细胞计数、拟胚体计数和ES细胞集落边缘细胞分化状态比较ES细胞在三种培养基中生长和分化差异。结果 :与BRL组比较 ,MEF组和BRL +MEF组细胞生长较快(P <0 .0 1) ,ES细胞集落边缘分化细胞较少 ;MEF组和BRL +MEF组无明显差异。结论 :在复苏后早期阶段ES细胞培养中 ,不宜单独应用BRL条件培养基 ,须用MEF饲养层或合并应用BRL条件培养基和MEF饲养层更多还原

【Abstract】 Aim: To study the method of using BRL conditioned medium in ES cells culture. Methods: ES cells were cultured in BRL conditioned medium,MEF feeder layer,and BRL conditioned medium plus MEF feeder layer respectively. Both the numbers of ES cells and embryonic bodies and the differentiated cells around the ES cell colonies at certain stages were studied to find the differences of growing speed of ES cells in three culture mediums. Results: On promoting the growth and maintaining the undifferentiated state of ES cells at the early stage after recovery,the effects of BRL conditioned medium were less than those of MEF feeder layer and those of BRL conditioned medium plus MEF feeder layer ( P <0.01),and there was no difference between MEF feeder layer and BRL conditioned medium plus MEF feeder layer ( P >0.05). Conclusion: ES cells should be cultivated on MEF feeder layer or in BRL conditioned medium plus MEF feeder layer at the early stage after recovering.更多还原