Generation of Human-Induced Pluripotent Stem Cells from Gut Mesentery-Derived Cells by Ectopic Expression of OCT4/SOX2/NANOG

Peking University Stem Cell Research Center, Beijing, People's Republic of China.
Cellular reprogramming 06/2010; 12(3):237-47. DOI: 10.1089/cell.2009.0103
Source: PubMed


Induced pluripotent stem (iPS) cells have been generated from human somatic cells by ectopic expression of defined transcription factors. Application of this approach in human cells may have enormous potential to generate patient-specific pluripotent stem cells. However, traditional methods of reprogramming in human somatic cells involve the use of oncogenes c-MYC and KLF4, which are not applicable to clinical translation. In the present study, we investigated whether human fetal gut mesentery-derived cells (hGMDCs) could be successfully reprogrammed into induced pluripotent stem (iPS) cells by OCT4, SOX2, and NANOG alone. We used lentiviruses to express OCT4, SOX2, NANOG, in hGMDCs, then generated iPS cells that were identified by morphology, presence of pluripotency markers, global gene expression profile, DNA methylation status, capacity to form embryoid bodies (EBs), and terotoma formation. iPS cells resulting from hGMDCs were similar to human embryonic stem (ES) cells in morphology, proliferation, surface markers, gene expression, and epigenetic status of pluripotent cell-specific genes. Furthermore, these cells were able to differentiate into cell types of all three germ layers both in vitro and in vivo, as shown by EB and teratoma formation assays. DNA fingerprinting showed that the human iPS cells were derived from the donor cells, and are not a result of contamination. Our results provide proof that hGMDCs can be reprogrammed into pluripotent cells by ectopic expression of three factors (OCT4, SOX2, and NANOG) without the use of oncogenes c-MYC and KLF4.

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    • "While OCT4, a member of the POU (Pit-Oct-Unc) transcription factor family, is essential for the maintenance of self-renewal capacities, NANOG (a downstream target of OCT4) contributes to the cell fate determination of pluripotent cells during embryogenesis [36,37]. OCT4 and NANOG are among the few key factors that enable the reprogramming of adult somatic cells into pluripotent stem cells [38-42]. Interestingly, induced pluripotent cells (iPSCs) generated from sheep fibroblasts have recently been demonstrated to exhibit an embryonic stem cell-like morphology and to express OCT4 and NANOG among other intracellular and surface markers associated with undifferentiated cells, as previously demonstrated in humans and mice [43]. "
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    • "The difficulties associated with the use of embryonic stem cells can be overcome by the use of induced pluripotent stem (iPS) cells. iPS cells were originally derived from adult fibroblasts by the introduction and expression of four genes, namely OCT3/4, SOX2, KLF4 and c-MYC [188]. Since iPS cells derived by reprogramming adult cells using the oncogenes KLF4 and c-MYC, which when expressed can lead to the development of teratomas, a number of investigators have been successful at reprogramming adult cells into iPS cells using other gene sets [189, 190]. "
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    • "EXPRESSION DURING LIMB REGENERATION Limb regeneration in Xenopus involves numerous signaling pathways (Beck et al., 2009), including BMP (Beck et al., 2006), Fgf (Yokoyama et al., 2000), and Wnt (Kawakami et al., 2006). Regulation of Sall4 expression during reprogramming and maintenance of pluripotency in ESC and somatic stem cells and probably during patterning also involves numerous context-dependent signaling pathways (Li et al., 2010). For example, in Drosophila melanogaster spalt/Sal expression is regulated during embryogenesis by Shh, dpp (BMP), and EGF signaling pathways, depending on the context (de Celis and Barrio, 2009). "
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