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当前位置: 首页 > 文章 > 影响山羊类胚胎干细胞分离克隆的因素分析 黑龙江畜牧兽医 2007 (4) 17-20
Position: Home > Articles > Analysis of influenctial factors on isolatin and clonging of goat embryonic stem-like cells Heilongjiang Animal Science and Veterinary Medicine 2007 (4) 17-20

影响山羊类胚胎干细胞分离克隆的因素分析

作  者：
杨丽芬;王清华;韩芬霞;项智锋;张晓建;王天友;常新耀
单  位：
河南科技学院动物科学学院
关键词：
山羊;原始生殖细胞;类ES细胞
摘  要：
将本地槐山羊胎儿的原始生殖细胞(PGCs)与其生殖嵴周围组织细胞共同分离,经传代培养后获得了具有干细胞特征的山羊类ES细胞。结果表明高糖DMEM培养基和低糖DMEM培养基相比较,低糖DMEM更适宜于山羊类ES细胞的分离与克隆;类ES细胞在山羊胎儿成纤维细胞饲养层上生长效果较好,可传4代或5代,而在小鼠成纤维细胞饲养层上,类ES细胞仅传3代;联合添加白血病抑制因子(LIF)、干细胞因子(SCF)和碱性成纤维细胞生长因子(bFGF)能显著提高山羊类ES细胞分离与克隆的效率;胎龄为30~45d的胎儿原代培养时可获得大量的细胞集落,克隆培养可传至5代,适合作山羊类ES细胞的分离培养。
译  名：
Analysis of influenctial factors on isolatin and clonging of goat embryonic stem-like cells
作  者：
YANG Li-fen,WANG Qing-hua,HAN Fen-xia,et al(Animal Science College of Henan Science and Technology Institute,Xinxiang 453003,China)
关键词：
Goat; primordial germ cells; embryonic stem-like cells
摘  要：
To study effects of culture media, feeder layer, growth factor and fetus age on isolation and passage of embryonic stem(ES) cell, primordial germ cells (PGCs) along with jacent gonadal anlage were isolated and co-cultured, from which ES-like cells were obtained. 5th passage was obtained by culturing PGCs of 30～45 d fetus, which can bring quantity of cell centralization. Compared with 3rd passages of DMEM of high glucose, 4th or 5th were obtained on low glucose DMEM. It can significantly increased its efficiency by adding leukaemia inhibitory factor(LIF), stem cell factor(SCF) and basic fibroblast growth factor (bFGF). In a word, 30～45 d fetus are suitable to isolate PGCs, DMEM (low glucose), goat embryonic fibroblasts feeder layer and addition of LIF+SCF+bFGF are advantageous condition for isolating and culturing of goat ES-like cells.

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