Generation of human androgenetic induced pluripotent stem cells

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. The contribution of both the maternal and paternal genomes is required for normal development. The maternal and paternal genetic contributions are important in early embryonic development and placental development, respectively 1,2. A uniparental embryo with two maternal genomes is termed parthenogenetic, whereas that with two paternal genomes is termed androgenetic. Uniparental embryos fail to develop 3,4. Recent studies have reported the derivation of human parthenogenetic embryonic stem cells (PgESCs) from parthenogenetic embryos 5-8. These studies have determined the methylation status and expression levels of imprinted genes in human PgESCs. Human androgenetic embryonic stem cells (AgESCs) can be generated from androgenetic embryos 9. Human AgESCs have androgenetic imprinting status because of the lack of the maternal genome. Bisulfite sequencing analysis of the methylation status of human AgESCs revealed that paternally imprinted genes (SNRPN and KCNQ1) were hypomethylated, whereas maternally imprinted genes (H19 and MEG3) were hypermethylated 9. These findings indicate that the androgenetic identity of uniparental embryonic stem cells (ESCs) can be confirmed by determining the methylation status of imprinted genes. Although human AgESCs are considered attractive cell lines, their generation poses ethical issues because of the use of human eggs and the destruction of human embryos. Although they are rare, parthenogenesis and androgenesis occur naturally in humans. Mature cystic ovarian teratomas, also known as dermoid cysts, arise from parthenogenetic activation of oocytes 10,11. Hydatidiform moles are abnormal pregnancies and can be classified as complete hydatidiform moles (CHM) or partial hydatidiform moles (PHM) depending on their genetic origin 12,13. Most CHM have an entirely paternal origin and lack

Stem cells research prospects towards precision medicine

“…Furthermore, human androgenetically induced pluripotent stem cells (AgHiP-SCs) were recently produced in research on primary androgenetic fi broblasts. AgHiPSCs can also be employed in regenerative medicine, genomic imprinting studies, imprinting-related development research, and human sickness modelling (37).…”