畜牧兽医学报2011，Vol.42Issue(8)：1187-1192,6.
山羊类ES细胞的分离与培养
Isolation and Culture of Goat ES-like Cells
摘要
Abstract
The aim of this study was to optimize and set up better goat feeder layer cell, culture medium, cell factors and passaging method for better growth of goat ES-like cells. Blastocysts obtained by whole embryo cultured, trypsin digestion and mechanical dissociation after enzymatic treatment from Wuhan goats 6-7 days after mating were cultivated. The effect of different cell layers, different cell factors in culture medium and different passaging methods on the cell survival and differentiation were compared. Alkaline phosphataseassay (ALP) and immunohistochemi-cal staining method were used to detect the activity of AKT and the expression of ES cells specific transcription factor including SSEA-1 and Oct-4, and further study the differentiation ability of goat ES-like cells. Significant differences are not noted between cell types that C2C12 can promote adherence(100%), better than MEF(81. 8%)and GEF(88. 5%), but not benefit for restraining undifferentiation state; the division of inner cell mass with high concentration of LIF and SCF cell factors could help the cells passage in undifferential condition (8 passages) ; the media contai-ning LIF and SCF, while adding heparin and insulin, was more conducive to cells passage (9 passages) ; the ESCs expressed the protein of SSEA-1 and Oct-4, in vitro these cells could differentiate into epithelial-like cells, vacuole-like cells and cardiomyocytes. The goat ES-like cells differentiating into variety cell types were obtained successfully.关键词
胚胎干细胞/内细胞团/山羊Key words
embryonic stem cell/ inner mass cell/ goat引用本文复制引用
刘艳,高其双,黄海军,邓兵,蒋思文..山羊类ES细胞的分离与培养[J].畜牧兽医学报,2011,42(8):1187-1192,6.基金项目
国家“863”计划(2007AA100505) （2007AA100505）
