TY - JOUR
T1 - Establishment of leukemia inhibitory factor (LIF)-independent iPS cells with potentiated Oct4
AU - Hirai, Hiroyuki
AU - Firpo, Meri
AU - Kikyo, Nobuaki
N1 - Funding Information:
We thank Toshio Kitamura for Plat-E cells and pMXs-IP retroviral vectors, Wei-Shou Hu and Andrew Yongky for microarray analyses, and Peter Karian for mouse experiments. mRNA microarray analyses were done at the University of Minnesota Genomics Center. Histology was performed at the University of Minnesota Masonic Cancer Center Comparative Pathology Shared Resource with support from the NIH Grant P30 CA077598. M.F. and N.K were supported by Richard M. Schulze Family Foundation . N.K. was supported by the NIH ( R01 GM098294 and R21 CA187232 ); Engdahl Family Foundation ; Grain-in-Aid of Research, Artistry and Scholarship, University of Minnesota ( 22802 ); and the University of Minnesota Foundation ( 4160-9227-13 ).
Publisher Copyright:
© 2015 The Authors.
PY - 2015/11/1
Y1 - 2015/11/1
N2 - Leukemia inhibitory factor (LIF) is widely used to establish and maintain naïve pluripotent stem cells, including mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). Although the combination of chemical inhibitors called 2i can establish mouse iPSCs without LIF from primed pluripotent stem cells, it has been difficult, if not impossible, to establish mouse iPSCs from differentiated somatic cells without LIF. We previously showed that the fusion gene of the transactivation domain of MyoD and the full-length Oct4 (M3O) increases the efficiency of making iPSCs when transduced into fibroblasts along with Sox2, Klf4, and c-Myc (M3O-SKM). Here, we report that M3O-SKM allows for establishment of iPSCs without exogenous LIF from mouse embryonic fibroblasts. The established iPSCs remained undifferentiated and maintained pluripotency over 90days without LIF as long as M3O was expressed. The iPSCs upregulated miR-205-5p, which was potentially involved in the LIF-independence by suppressing the two signaling pathways inhibited by 2i. The result indicates that potentiated Oct4 can substitute for the LIF signaling pathway, providing a novel model to link Oct4 and LIF, two of the most significant players in naïve pluripotency.
AB - Leukemia inhibitory factor (LIF) is widely used to establish and maintain naïve pluripotent stem cells, including mouse embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). Although the combination of chemical inhibitors called 2i can establish mouse iPSCs without LIF from primed pluripotent stem cells, it has been difficult, if not impossible, to establish mouse iPSCs from differentiated somatic cells without LIF. We previously showed that the fusion gene of the transactivation domain of MyoD and the full-length Oct4 (M3O) increases the efficiency of making iPSCs when transduced into fibroblasts along with Sox2, Klf4, and c-Myc (M3O-SKM). Here, we report that M3O-SKM allows for establishment of iPSCs without exogenous LIF from mouse embryonic fibroblasts. The established iPSCs remained undifferentiated and maintained pluripotency over 90days without LIF as long as M3O was expressed. The iPSCs upregulated miR-205-5p, which was potentially involved in the LIF-independence by suppressing the two signaling pathways inhibited by 2i. The result indicates that potentiated Oct4 can substitute for the LIF signaling pathway, providing a novel model to link Oct4 and LIF, two of the most significant players in naïve pluripotency.
KW - Induced pluripotent stem cells
KW - Leukemia inhibitory factor
KW - Pluripotency
KW - miRNA
UR - http://www.scopus.com/inward/record.url?scp=84942304239&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84942304239&partnerID=8YFLogxK
U2 - 10.1016/j.scr.2015.09.002
DO - 10.1016/j.scr.2015.09.002
M3 - Article
C2 - 26413786
AN - SCOPUS:84942304239
SN - 1873-5061
VL - 15
SP - 469
EP - 480
JO - Stem Cell Research
JF - Stem Cell Research
IS - 3
ER -