Tanaka Tetsuya S
Department of Biological Sciences, Chemical and Biomolecular Engineering, University of Notre Dame, 49 Galvin Life Sciences, Notre Dame, IN, 46556, USA.
Methods Mol Biol. 2016;1341:295-319. doi: 10.1007/7651_2015_228.
This chapter describes standard techniques to (1) maintain mouse embryonic stem cell culture, (2) deliver transgenes into mouse embryonic stem cells mediated by electroporation, nucleofection, lipofection, and retro/lentiviruses, and (3) assess the pluripotency of mouse embryonic stem cells. The last part of this chapter presents induction of random cell differentiation followed by the alkaline phosphatase and embryoid body formation assays, immunofluorescence microscopy, and the teratoma formation assay.
(1)维持小鼠胚胎干细胞培养;(2)通过电穿孔、核转染、脂质体转染以及逆转录病毒/慢病毒介导将转基因导入小鼠胚胎干细胞;(3)评估小鼠胚胎干细胞的多能性。本章最后一部分介绍了诱导随机细胞分化,随后进行碱性磷酸酶和胚状体形成分析、免疫荧光显微镜检查以及畸胎瘤形成分析。