Improved genetic manipulation of human embryonic stem cells
Improved genetic manipulation of human embryonic stem cells
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DOI：
10.1038/nmeth.1200
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发表时间：
2008-05-01
期刊：
影响因子：
48
通讯作者：
Mummery, Christine L.
中科院分区：
文献类型：
--
作者：
Braam, Stefan R.;Denning, Chris;Mummery, Christine L.
低效率的转染限制了遗传操作人胚胎干细胞（hESC）的能力，并且细胞衍生和培养方法的差异需要优化转染方案。我们将具有不同生长要求的多个独立的hESC系瞬时转移到标准化的无饲养层培养中，并优化了克隆生长和有效基因转移的条件，而不丧失多能性。稳定转染的细胞系保持分化潜能，大多数细胞系显示正常核型。
Low efficiency of transfection limits the ability to genetically manipulate human embryonic stem cells (hESCs), and differences in cell derivation and culture methods require optimization of transfection protocols. We transiently transferred multiple independent hESC lines with different growth requirements to standardized feeder-free culture, and optimized conditions for clonal growth and efficient gene transfer without loss of pluripotency. Stably transfected lines retained differentiation potential, and most lines displayed normal karyotypes.