Derivation of a Homozygous Human Androgenetic Embryonic Stem Cell Line

Abstract

Human embryonic stem cells (hESCs) have long been considered as a promising source for cell replacement therapy. However, one major obstacle for the use of these cells is immune compatibility. Histocompatible human parthenogenetic ESCs have been reported as a new method for generating human leukocyte antigen (HLA)-matched hESCs. To further investigate the possibility of obtaining histocompatible stem cells from uniparental embryos, we tried to produce androgenetic haploid human embryos by injecting a single spermatozoon into enucleated human oocyte, and establish human androgenetic embryonic stem (hAGES) cell lines from androgenetic embryos. In the present study, a diploid hAGES cell line has been established, which exhibits typical features of human ESCs, including the expression of pluripotency markers, having differentiation potential in vitro and in vivo, and stable propagation in an undifferentiated state (>P40). Bisulfite sequencing of the H19, Snrpn, Meg3, and Kv imprinting control regions suggested that hAGES cells maintained to a certain extent a sperm methylation pattern. Genome-wide single nucleotide polymorphism, short tandem repeat, and HLA analyses revealed that the hAGES cell genome was highly homozygous. These results suggest that hAGES cells from spermatozoon could serve as a useful tool for studying the mechanisms underlying genomic imprinting in humans. It might also be used as a potential resource for cell replacement therapy as parthenogenetic stem cells.

Generation of human androgenetic induced pluripotent stem cells

“…Our SNP analysis showed 99.53% homozygosity in AgFibs, 99.48% in AgHiPSCs, and 72.72% in H9 cells. According Ding et al 9 , human AgESCs were 99.39% homozygous, in contrast to 72.85% homozygosity in H9 cells. These data confirmed that our androgenetic cell lines were homogeneous.…”

Generation and application of mammalian haploid embryonic stem cells

“… successfully generated haESCs from Macaca fascicularis monkey parthenotes. Nevertheless, preliminary attempts to derive human haESCs resulted in a homozygous diploid AG ESC line ; thus whether haESCs can be derived from humans remains an open question.…”

“…Recently, by combined application of a modified single-cell enzymatic dissociation method [37][38][39] and regular cell sorting, Yang et al [40] successfully generated haESCs from Macaca fascicularis monkey parthenotes. Nevertheless, preliminary attempts to derive human haESCs resulted in a homozygous diploid AG ESC line [41]; thus whether haESCs can be derived from humans remains an open question.…”

Derivation and application of pluripotent stem cells for regenerative medicine

“…In addition, cynomolgus monkey ESCs with higher pluripotency with which monkey chimeric fetuses were generated have been obtained . Human parthenogenetic ESCs Shuai et al, 2015) and homozygous human androgenetic ESCs (Ding et al, 2015) were derived, providing important tools for future therapeutic use in a clinical setting.…”