小鼠胎肺Ⅲ型上皮细胞的分离纯化及原代培养Isolation,Purification and Primary Culture of Type Ⅱ Alveolar Epithelial Cells from Fetal Mouse
张明凤;张彦定;王昕;王虹;张龙斌;
摘要(Abstract):
用免疫黏附和差速离心法分离和纯化小鼠胎肺Ⅱ型细胞,所得细胞纯度达(90±2)%,产量为每5只小鼠胎肺收获(23.6±7)×106个细胞.原代培养的小鼠胎肺Ⅱ型细胞在1224 h开始贴壁,在3624 h开始贴壁,在3648 h呈指数生长,72 h后生长减缓,448 h呈指数生长,72 h后生长减缓,45 d左右细胞开始变形分化.免疫荧光鉴定胎肺Ⅱ型细胞表面活性蛋白C(SP-C)染色阳性,透射电镜观察到细胞内板层小体,细胞中proSP-C蛋白在培养48 h后表达较高.体外病毒转染成功,可见强烈荧光表达.成功建立了小鼠胎肺Ⅱ型上皮细胞的培养模型.
关键词(KeyWords): 胎鼠;肺泡Ⅱ型细胞;分离;鉴定;原代培养
基金项目(Foundation): 国家自然科学基金资助项目(30771132);; 福建省青年科技人才创新项目(2007F3024);; 福建省自然科学基金资助项目(2009J01123)
作者(Authors): 张明凤;张彦定;王昕;王虹;张龙斌;
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