A Murine ESC-like State Facilitates Transgenesis and Homologous Recombination in Human Pluripotent Stem Cells

被引：181

作者：

Buecker, Christa ^{[1
,4
]}

Chen, Hsu-Hsin ^{[1
,4
]}

Polo, Jose Maria ^{[1
,2
,4
]}

Daheron, Laurence ^{[1
,4
]}

Bu, Lei ^{[3
,4
]}

Barakat, Tahsin Stefan ^{[6
]}

Okwieka, Patricia ^{[1
]}

Porter, Andrew ^{[7
]}

Gribnau, Joost ^{[6
]}

Hochedlinger, Konrad ^{[1
,2
,4
,5
]}

Geijsen, Niels ^{[1
,4
,8
]}

机构：

[1] Harvard Univ, Massachusetts Gen Hosp, Sch Med, Ctr Regenerat Med, Boston, MA 02114 USA

[2] Harvard Univ, Massachusetts Gen Hosp, Sch Med, Ctr Canc, Boston, MA 02114 USA

[3] Harvard Univ, Massachusetts Gen Hosp, Sch Med, Cardiovasc Res Ctr, Boston, MA 02114 USA

[4] Harvard Univ, Harvard Stem Cell Inst, Cambridge, MA 02138 USA

[5] Harvard Univ, Dept Stem Cell & Regenerat Biol, Cambridge, MA 02138 USA

[6] Univ Med Ctr, Erasmus MC, Dept Reprod & Dev, NL-3000 CA Rotterdam, Netherlands

[7] Univ London Imperial Coll Sci Technol & Med, Gene Therapy Grp, Dept Haematol, London W12 0NN, England

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

来源：

CELL STEM CELL | 2010年 / 6卷 / 06期

关键词：

HUMAN BLASTOCYSTS; MOUSE EMBRYOS; GROUND-STATE; SELF-RENEWAL; METHYLATION; CHROMATIN; EPIBLAST; LINES; DIFFERENTIATION; GENERATION;

D O I：

10.1016/j.stem.2010.05.003

中图分类号：

Q813 [细胞工程];

学科分类号：

摘要：

Murine pluripotent stem cells can exist in two functionally distinct states, LIF-dependent embryonic stem cells (ESCs) and bFGF-dependent epiblast stem cells (EpiSCs). However, human pluripotent cells so far seemed to assume only an epiblast-like state. Here we demonstrate that human iPSC reprogramming in the presence of LIF yields human stem cells that display morphological, molecular, and functional properties of murine ESCs. We termed these hLR5 iPSCs because they require the expression of five ectopic reprogramming factors, Oct4, Sox2, Klf4, cMyc, and Nanog, to maintain this more naive state. The cells are "metastable" and upon ectopic factor withdrawal they revert to standard human iPSCs. Finally, we demonstrate that the hLR5 state facilitates gene targeting, and as such provides a powerful tool for the generation of recombinant human pluripotent stem cell lines.

引用

页码：535 / 546

页数：12

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