Abstract
Somatic cells can be reprogrammed into induced pluripotent stem (iPS) cells by the transcription factors Oct4, Sox2, and Klf4 in combination with c-Myc. Recently, Sox2 plus Oct4 was shown to reprogram fibroblasts and Oct4 alone was able to reprogram mouse and human neural stem cells (NSCs) into iPS cells. Here, we report that Bmi1 leads to the transdifferentiation of mouse fibroblasts into NSC-like cells, and, in combination with Oct4, can replace Sox2, Klf4 and c-Myc during the reprogramming of fibroblasts into iPS cells. Furthermore, activation of sonic hedgehog signaling (by Shh, purmorphamine, or oxysterol) compensates for the effects of Bmi1, and, in combination with Oct4, reprograms mouse embryonic and adult fibroblasts into iPS cells. One- and two-factor iPS cells are similar to mouse embryonic stem cells in their global gene expression profile, epigenetic status, and in vitro and in vivo differentiation into all three germ layers, as well as teratoma formation and germline transmission in vivo. These data support that converting fibroblasts with Bmi1 or activation of the sonic hedgehog pathway to an intermediate cell type that expresses Sox2, Klf4, and N-Myc allows iPS generation via the addition of Oct4.
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Acknowledgements
We are grateful to Drs Goberdhan P Dimri (NorthShore University HealthSystem Research Institute, USA)and Gou Young Koh (Korea Advanced Institute of Science and Technology, Republic of Korea) for kindly providing the pBabe-Bmi1, pMX-Oct4, pMX-Sox2, pMX-Klf4, and pMX-C-Myc constructs. We thank Jihyun Kim, Jihye Hwang, and Suhyun Kwon for technical support. This research was supported by a grant (SC-5150) from the Stem Cell Research Center of the 21st Century Frontier Research Program funded by the Ministry of Education, Science and Technology, Republic of Korea, a grant (09172KFDA653) from the Korea Food and Drug Administration, and a grant (2010-0020347) from National Research Foundation (NRF) funded by the Korea government (MEST). Work in Germany was supported by the Max Planck Society and the Federal Ministry of Education and Research (BMBF) on Cell-Based Regenerative Medicine (Grant 01GN0539).
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( Supplementary information is linked to the online version of the paper on the Cell Research website.)
Supplementary information
Supplementary information, Figure S1
Phase-contrast photographs of cell morphology and immunofluoresence of reprogrammed iPS cells. (PDF 176 kb)
Supplementary information, Figure S2
Characterization of BO-iPS cells. (PDF 135 kb)
Supplementary information, Figure S3
Generation and characterization of BO-iPS clones (1–6) from established BO-iPS cells by single cell culture. (PDF 189 kb)
Supplementary information, Figure S4
Characterization of ShO-iPS generated from MEFs by treatment of Shh and retroviral transduction with Oct4. (PDF 157 kb)
Supplementary information, Figure S5
Generation and characterization of PO-iPS cells and OxyO-iPS cells. (PDF 222 kb)
Supplementary information, Figure S6
Generation and characterization of 1F PO-iPS cells. (PDF 203 kb)
Supplementary information, Figure S7
Generation and characterization of 1F OxyO-iPS-TTF cells. (PDF 189 kb)
Supplementary information, Figure S8
Global gene expression analyses by DNA microarray (PDF 113 kb)
Supplementary information, Figure S9
Karyotype analysis of iPS cells. (PDF 105 kb)
Supplementary information, Table S1
Population doubling time (PDF 35 kb)
Supplementary information, Table S2
Genes belonging to groups in Supplementary information, Figure S8 (PDF 99 kb)
Supplementary information, Table S3
Primers used in this study (PDF 134 kb)
Supplementary information, Data S1
Supplemental Methods (PDF 63 kb)
Supplementary information, Table S4
Antibodies and dilutions used in this study (PDF 48 kb)
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Moon, JH., Heo, J., Kim, J. et al. Reprogramming fibroblasts into induced pluripotent stem cells with Bmi1. Cell Res 21, 1305–1315 (2011). https://doi.org/10.1038/cr.2011.107
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DOI: https://doi.org/10.1038/cr.2011.107
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