Improved genetic manipulation of human embryonic stem cells

被引：63

作者：

Braam, Stefan R. ^{[1
,2
]}

Denning, Chris ^{[3
]}

van den Brink, Stieneke ^{[1
]}

Kats, Peter ^{[4
]}

Hochstenbach, Ron ^{[4
]}

Passier, Robert ^{[1
,2
]}

Mummery, Christine L. ^{[1
,2
,3
,5
]}

机构：

[1] Hubrecht Inst Dev Biol & Stem Cell Res, NL-3584 CT Utrecht, Netherlands

[2] Leiden Univ, Med Ctr, Dept Anat & Embryol, NL-2300 RC Leiden, Netherlands

[3] Univ Nottingham, Ctr Biomol Sci, Wolfson Ctr Stem Cells Tissue Engn & Modelling ST, Nottingham NG7 2RD, England

[4] Univ Med Ctr Utrecht, Dept Biomed Genet, NL-3508 AB Utrecht, Netherlands

[5] Interuniv Cardiol Inst Netherlands, NL-3511 Utrecht, Netherlands

来源：

NATURE METHODS | 2008年 / 5卷 / 05期

基金：

英国生物技术与生命科学研究理事会; 英国医学研究理事会;

关键词：

D O I：

10.1038/NMETH.1200

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：389 / 392

页数：4

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