Derivation of a Homozygous Human Androgenetic Embryonic Stem Cell Line

Stem Cells Dev. 2015 Oct 1;24(19):2307-16. doi: 10.1089/scd.2015.0031. Epub 2015 Aug 24.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Androgens / metabolism
  • Cell Line
  • Cells, Cultured
  • Diploidy
  • Embryo, Mammalian / cytology*
  • Embryo, Mammalian / metabolism
  • Embryonic Stem Cells / cytology*
  • Embryonic Stem Cells / metabolism
  • Female
  • Gene Expression
  • Haploidy
  • Homeodomain Proteins / genetics
  • Homozygote
  • Humans
  • In Situ Hybridization, Fluorescence
  • Male
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3 / genetics
  • Oocytes / cytology*
  • Oocytes / metabolism
  • RNA, Long Noncoding / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • SOXB1 Transcription Factors / genetics
  • Sperm Injections, Intracytoplasmic
  • Spermatozoa / cytology*
  • Spermatozoa / metabolism
  • Spermatozoa / transplantation
  • snRNP Core Proteins / genetics

Substances

  • Androgens
  • H19 long non-coding RNA
  • Homeodomain Proteins
  • NANOG protein, human
  • Nanog Homeobox Protein
  • Octamer Transcription Factor-3
  • POU5F1 protein, human
  • RNA, Long Noncoding
  • SOX2 protein, human
  • SOXB1 Transcription Factors
  • snRNP Core Proteins