Afanassieff Marielle, Osteil Pierre, Savatier Pierre
INSERM, U846, 18 Avenue du Doyen Lépine, F-69500, Bron, France.
Stem Cell and Brain Research Institute, F-69500, Bron, France.
Methods Mol Biol. 2016;1341:49-66. doi: 10.1007/7651_2015_209.
Here we have described a procedure to generate embryonic stem cell (ESC) lines from rabbit preimplantation blastocysts. We have also provided detailed procedures to characterize the resulting ESC lines, such as the analysis of pluripotency marker expression by reverse transcription quantitative polymerase chain reaction, immunolabeling, and fluorescence-associated cell sorting; evaluation of pluripotency by teratoma production; and assessment of genetic stability by karyotyping.
在此,我们描述了一种从兔植入前囊胚中生成胚胎干细胞(ESC)系的方法。我们还提供了详细的程序来鉴定所产生的ESC系,例如通过逆转录定量聚合酶链反应、免疫标记和荧光相关细胞分选分析多能性标志物的表达;通过畸胎瘤形成评估多能性;以及通过核型分析评估遗传稳定性。