C57BL/6J×129/J杂交小鼠ES细胞系的建株

CHINESE JOURNAL OF PATHOPHYSIOLOGY(2006)

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Abstract
目的:建立C57BL/6J×129/J杂交小鼠ES细胞系.方法:收集3.5 d.p.c.的囊胚,培养在预先铺有小鼠成纤维细胞(MEFs)的高糖DMEM培养液中.3-4 d后,挑出内细胞团(ICM),消化后重新种到新鲜的有MEFS培养液中.等到有典型的ES样集落长出,即传代以得到永久ES细胞系.通过分析碱性磷酸酶活性,SSEA-1,Oct-4的表达和形成畸胎瘤的能力来鉴定ES细胞的多向分化能力.结果:获得的两个C57BL/6J×129/J杂交小鼠ES细胞系绝大多数细胞具有正常的核型(40,XY),碱性磷酸酶染色阳性,SSEA-1,Oct-4表达阳性,ES细胞注入SCID鼠后可获得来自3个胚层的组织.结论:建立了两株具有长期自我更新能力和多向分化潜能的C57BL/6J×129/J杂交小鼠ES细胞系.
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Key words
Mice,Embryonic stem cells,Hybridization
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