Generation of Naive Induced Pluripotent Stem Cells from Rhesus Monkey Fibroblasts

Abstract

Conventional embryonic stem cells (ESCs) or induced pluripotent stem cells (iPSCs) derived from primates resemble mouse epiblast stem cells, raising an intriguing question regarding whether the naive pluripotent state resembling mouse embryonic stem cells (mESCs) exists in primates and how to capture it in vitro. Here we identified several specific signaling modulators that are sufficient to generate rhesus monkey fibroblast-derived iPSCs with the features of naive pluripotency in terms of growth properties, gene expression profiles, self-renewal signaling, X-reactivation, and the potential to generate cross-species chimeric embryos. Interestingly, together with recent reports of naive human pluripotent stem cells, our findings suggest several conserved signaling pathways shared with rodents and specific to primates, providing significant insights for acquiring naive pluripotency from other species. In addition, the derivation of rhesus monkey naive iPSCs also provides a valuable cell source for use in preclinical research and disease modeling.

Discrimination of Stem Cell Status after Subjecting Cynomolgus Monkey Pluripotent Stem Cells to Naïve Conversion

“…To examine whether our naïve-like ESCs would contribute to interspecific chimeras as shown by Fang et al 18,. they were injected into mouse pre-implantation embryos.…”

“…To mitigate the ethical concerns in using human PSCs, translational research using non-human primates might provide precise indicators for human PSC application. Naïve-like conversion of non-human primate ESCs17 and iPSCs18 has been demonstrated. Both types of naïve-like converted PSCs could contribute to chimeric embryos but only in minor ways via unknown species-specific features despite the successful demonstration of some naïve characteristics.…”

Efficient Generation of Chemically Induced Mesenchymal Stem Cells from Human Dermal Fibroblasts

“…NHSM can promote the dedifferentiation of human pluripotent stem cells from the ground stage to the naïve stage4546, and requires at least 9 factors (LIF, TGFβ1, bFGF, insulin, 2-mercaptoethanol, PD0325901, CHIR99021, SP600125, and SB203580), while the use of 11 factors (LIF, TGFβ1, bFGF, insulin, 2-mercaptoethanol, Y27632, Go6983, PD0325901, CHIR99021, SP600125, and SB203580) might have the best efficiency45. Since both NHSM and our cocktail support dedifferentiation, there may be some overlapping signals that trigger the conversion of cell state.…”

Contributions of Mammalian Chimeras to Pluripotent Stem Cell Research

“…Indeed, the rat-mouse species pairing has been recently used to demonstrate organ-specific contribution in interspecies chimeras (Usui et al., 2012, Kobayashi et al., 2010, Isotani et al., 2011). Monkey iPSCs, when injected into mouse preimplantation embryos, were non-chimeric at mid-gestation fetal stages (Fang et al., 2014). However, naive monkey ESCs were able to form fetal chimeras (14.3%) and preimplantation chimeric outgrowths in vitro (51.8% at 2 DIV), (Chen et al., 2015), for a chimeric index of 3.62, which is likely artificially inflated due to the short period of in vitro growth.…”