Abstract
Pluripotent stem cells can be established from parthenogenetic embryos, which only possess maternal alleles with maternal-specific imprinting patterns. Previously, we and others showed that parthenogenetic embryonic stem cells (pESCs) and parthenogenetic induced pluripotent stem cells (piPSCs) progressively lose the bimaternal imprinting patterns. As ESCs and iPSCs are naïve pluripotent stem cells, parthenogenetic primed pluripotent stem cells have not yet been established, and thus, their imprinting patterns have not been studied. Here, we first established parthenogenetic epiblast stem cells (pEpiSCs) from 7.5 dpc parthenogenetic implantation embryos and compared the expression patterns and DNA methylation status of the representative imprinted genes with biparental EpiSCs. We found that there were no striking differences between pEpiSCs and biparental EpiSCs with respect to morphology, pluripotency gene expression, and differentiation potential, but there were differences in the expression and DNA methylation status of imprinted genes (H19, Igf2, Peg1, and Peg3). Moreover, pEpiSCs displayed a different DNA methylation pattern compared with that of parthenogenetic neural stem cells (pNSCs), which showed a typical bimaternal imprinting pattern. These results suggest that both naïve pluripotent stem cells and primed pluripotent stem cells have an unstable imprinting status.
Highlights
The mammalian genome is inherited biparentally from oocytes and sperm
After culturing the unfertilized oocytes in CZB medium containing SrCl2 and cytochalasin B for 6 h, 79% (27/34) of the activated oocytes had developed into two-cell embryos and approximately 53% (18/34) of the parthenogenetic embryos progressed to the blastocyst stage (Figure 1a), which were transferred into pseudopregnant surrogate mice (2.5 dpc)
We hypothesized that there would be some abnormal phenotypes, such as poor proliferation rate and differentiation inefficiency, due to the unbalanced expression of imprinted genes; we could not observe any clear differences between parthenogenetic epiblast stem cells (pEpiSCs) and biparental epiblast stem cells (EpiSCs) with respect to the proliferation rate and the efficiency of differentiation
Summary
The mammalian genome is inherited biparentally from oocytes (maternal) and sperm (paternal). Naïve pluripotent stem cells, including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), are the in vivo counterparts of the blastocyst inner cell mass [11,12], while primed pluripotent stem cells, including epiblast stem cells (EpiSCs), are in a post-implantation epiblast-like state (late gastrula stage) [13,14,15] These two types of pluripotent stem cells are distinct in many aspects, such as differentiation potential, signaling pathways regulating their self-renewal, transcriptional and epigenetic profiles, mechanism of Oct regulation, energy metabolism, and chimera formation ability [16,17,18,19]
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