Abstract
In humans, parthenogenesis and androgenesis occur naturally in mature cystic ovarian teratomas and androgenetic complete hydatidiform moles (CHM), respectively. Our previous study has reported human parthenogenetic induced pluripotent stem cells from ovarian teratoma–derived fibroblasts and screening of imprinted genes using genome-wide DNA methylation analysis. However, due to the lack of the counterparts of uniparental cells, identification of new imprinted differentially methylated regions has been limited. CHM are inherited from only the paternal genome. In this study, we generated human androgenetic induced pluripotent stem cells (AgHiPSCs) from primary androgenetic fibroblasts derived from CHM. To investigate the pluripotency state of AgHiPSCs, we analyzed their cellular and molecular characteristics. We tested the DNA methylation status of imprinted genes using bisulfite sequencing and demonstrated the androgenetic identity of AgHiPSCs. AgHiPSCs might be an attractive alternative source of human androgenetic embryonic stem cells. Furthermore, AgHiPSCs can be used in regenerative medicine, for analysis of genomic imprinting, to study imprinting-related development, and for disease modeling in humans.
Highlights
In humans, parthenogenesis and androgenesis occur naturally in mature cystic ovarian teratomas and androgenetic complete hydatidiform moles (CHM), respectively
We report for the first time generation of human androgenetic induced pluripotent stem cells (AgHiPSCs) from CHM-derived fibroblasts
We isolated fibroblasts from a CHM resembling a bunch of grapes (Supplementary Data Fig. S1a) using collagenase/hyaluronidase to dissociate the tissue into single cells, as described for mouse and human mammary gland tissues[25,26]
Summary
Biparental fibroblasts showed somatic imprinting patterns in MEST (Supplementary Data Fig. S1c) These results confirm the androgenetic identity of AgFibs. We confirmed the expression of layer-specific gene markers in the endoderm (AFP, GATA4, and SOX17), mesoderm (MSX and BRACHYURY), and ectoderm (PAX6) of differentiated EBs by using RT-PCR analysis (Supplementary Data Fig. S7). These cells were positive for the markers of endoderm (AFP), mesoderm (NKX2.5 and CTNT), and ectoderm (TUJ1) in immunocytochemical analysis (Fig. 3a). Epigenetic instability of the imprinted genes H19 and MEST in human ESCs was previously reported[21,27,28,29] These results demonstrate the androgenetic imprinting status of AgHiPSCs
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