小鼠卵母细胞的孤雌激活与ES细胞样集落分离  

Mouse parthenogenesis and isolation of ES cell-likecolonies from parthenogenetic embryos

作　　者：曹鸿国[1] 刘俊平[1] 张涌[1] 

机构地区：[1]西北农林科技大学生物工程研究所,陕西杨凌712100

出　　处：《西北农林科技大学学报（自然科学版）》2005年第3期1-4,10,共5页Journal of Northwest A&F University(Natural Science Edition)

基　　金：国家"863"高技术项目(2001AA213081)

摘　　要：　对小鼠卵母细胞进行孤雌激活用于单性生殖研究。取成年昆明白小鼠进行超数排卵,乙醇激活,同小鼠输卵管上皮细胞共培养并添加mM16培养基培养至早期囊胚阶段,移至小鼠胎儿成纤维细胞饲养层上,换成ES细胞培养液继续培养,分离消化ICM,重新接种,对分离出的ES样细胞集落进行分离培养鉴定。结果显示,小鼠卵母细胞激活率为96.99%,2-细胞发育率为93.17%,桑椹胚发育率为80.33%,囊胚发育率为73.22%。小鼠孤雌激活囊胚中分离的ES样细胞集落具有一系列ES细胞所特有的特征,如呈现出岛屿状形态,碱性磷酸酶染色为阳性,体外能够自发分化成上皮样或单个散在分布的细胞等。试验证明,小鼠孤雌胚中可以分离出ES细胞样集落。In order to study parthenogenesis in depth,ES cells from parthenogenetic embryos of mice were isolated and cultured.Adult Kunming mice were superovulated with PMSG and HCG,and then oocytes were taken from mice oviducts.After treating 7 min with ethanol,oocytes were put on the feeder layer of mouse oviduct epidermal cells with mM16 media.When developed to the stage of early blastocyst,oocytes were transferred to the feeder layer of mouse embryo fibroblast with cardiomyocyte media conditioned,and then ICMs were digested and recultured.ES cell-like colonies were isolated after 2 days,identificated from morphology and with AKP staining and by differention in vitro.Results show that activation rate of mouse oocyte was 96.99%,2-cell development rate 93.17%,morula development rate 80.33%,and blastocyst development rate 73.22%.ES cell-like colonies were island-like,positive by AKP staining,and can spontaneously differentiate into epidermal-like cells or others around them.This demonstrates that ES cell-like colonies can be isolated from mice parthenogenetic embryos.

关 键 词：小鼠 卵母细胞 孤雌激活 ES细胞样集落 

分 类 号：Q813.7[生物学—生物工程]