Germline-competent mouse-induced pluripotent stem cell lines generated on human fibroblasts without exogenous leukemia inhibitory factor.
Induced pluripotent stem (iPS) cells have attracted enormous attention due to their vast potential in regenerative medicine, pharmaceutical screening and basic research. Most prior established iPS cell lines were derived and maintained on mouse embryonic fibroblast (MEF) cells supplemented with exogenous leukemia inhibitory factor (LIF). Drawbacks of MEF cells impede optimization as well as dissection of reprogramming events and limit the usage of iPS cell derivatives in therapeutic applications. In this study, we develop a reproducible protocol for efficient reprogramming mouse neural progenitor cells (NPCs) on human foreskin fibroblast (HFF) cells via retroviral transfer of human transcriptional factors OCT4/SOX2/KLF4/C-MYC. Two independent iPS cell lines are derived without exogenous LIF. They display typical undifferentiated morphology and express pluripotency markers Oct4 and Sox2. Transgenes are inactivated and the endogenous Oct4 promoter is completely demethylated in the established iPS cell lines, indicating a fully reprogrammed state. Moreover, the iPS cells can spontaneously differentiate or be induced into various cell types of three embryonic germ layers in vitro and in vivo when they are injected into immunodeficient mice for teratoma formation. Importantly, iPS cells extensively integrate with various host tissues and contribute to the germline when injected into the blastocysts. Interestingly, these two iPS cell lines, while both pluripotent, exhibit distinctive differentiation tendencies towards different lineages. Taken together, the data describe the first genuine mouse iPS cell lines generated on human feeder cells without exogenous LIF, providing a reliable tool for understanding the molecular mechanisms of nuclear reprogramming.
Li
Chunliang
C
Key Laboratory of Stem Cell Biology, Institute of Health Sciences, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences/Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Yu
Hongyao
H
Ma
Yu
Y
Shi
Guilai
G
Jiang
Jing
J
Gu
Junjie
J
Yang
Ying
Y
Jin
Shibo
S
Wei
Zhe
Z
Jiang
Hua
H
Li
Jinsong
J
Jin
Ying
Y
D000818:Animals; D005347:Fibroblasts / Q000166:cytology; D005854:Germ Cells; D006801:Humans; D000090062:Kruppel-Like Factor 4; D053605:Leukemia Inhibitory Factor / Q000494:pharmacology*; D051379:Mice; D039904:Pluripotent Stem Cells / Q000166:cytology*
C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D053605:Leukemia Inhibitory Factor
Nat Biotechnol. 2008 Aug;26(8):916-24
18594521
Cell. 2009 Feb 6;136(3):411-9
19203577
Cell. 2006 Aug 25;126(4):663-76
16904174
Nat Biotechnol. 2007 Oct;25(10):1177-81
17724450
Curr Biol. 2008 Jun 24;18(12):890-4
18501604
Hum Mol Genet. 2008 Sep 1;17(17):2654-64
18544626
Stem Cells. 2008 Oct;26(10):2467-74
18635867
Nature. 2008 May 22;453(7194):519-23
18497825
Science. 2008 Aug 1;321(5889):699-702
18276851
PLoS Biol. 2008 Oct 21;6(10):e253
18942890
Cell Stem Cell. 2007 Sep 13;1(3):245-7
18371358
Cell Stem Cell. 2008 Dec 4;3(6):595-605
19041776
Science. 2007 Dec 21;318(5858):1917-20
18029452
Nat Biotechnol. 2008 Nov;26(11):1276-84
18931654
Blood. 2009 May 28;113(22):5476-9
19299331
Cell. 2009 Mar 6;136(5):964-77
19269371
Cell Res. 2007 Sep;17(9):792-803
17768400
Cell Res. 2009 Feb;19(2):173-86
18982022
Cell Stem Cell. 2008 Sep 11;3(3):340-5
18786420
Science. 2008 Nov 7;322(5903):945-9
18818365
Nature. 2008 Jul 31;454(7204):646-50
18594515
Science. 2008 Nov 7;322(5903):949-53
18845712
Cell. 2008 Apr 18;133(2):250-64
18423197
Cell. 2007 Nov 30;131(5):861-72
18035408