Histocompatible Embryonic Stem Cells by Parthenogenesis

Abstract

Genetically matched pluripotent embryonic stem (ES) cells generated via nuclear transfer or parthenogenesis (pES cells) are a potential source of histocompatible cells and tissues for transplantation. After parthenogenetic activation of murine oocytes and interruption of meiosis I or II, we isolated and genotyped pES cells and characterized those that carried the full complement of major histocompatibility complex (MHC) antigens of the oocyte donor. Differentiated tissues from these pES cells engrafted in immunocompetent MHC-matched mouse recipients, demonstrating that selected pES cells can serve as a source of histocompatible tissues for transplantation.

A highly homozygous and parthenogenetic human embryonic stem cell line derived from a one-pronuclear oocyte following in vitro fertilization procedure

“…These cells could be used as autologous stem cells for the oocyte donor, but were of limited use for storage in the hESC bank owing to the heterozygosity of HLA loci. In mice, the incidence of heterozygosity of the major histocompatibility complex (MHC) is 33% in pESCs [33]. In recent studies in humans, all established pESCs were heterozygous in HLA loci [13,34].…”

“…In addition, 16 STR loci scattered in different regions of 12 different chromosomes, plus three HLA (-A, -B and -DRB) loci showing one allelic expression pattern and matching one allele of the maternal genome add further proof for the uniparental origin of this cell line. Genome-wide SNP analysis is regarded as a good tool for identifying the homozygous state of pESCs [33,34]. In this study, a high-resolution SNP microarray (500K) was used to identify the homozygosity of chHES-32 cells.…”

“…Thus, we conclude that chHES-32 cells are a homozygous pESC line. It is particularly interesting to compare the homozygous state of this cell line with the pESCs that are derived from artificially activated oocytes in mice [32,33] and in humans [13,34]. Artificial parthenogenesis is routinely induced by activating MII oocytes and preventing second polar body extrusion, which is thought to produce diploid embryos that are more likely to develop to the blastocyst stage [9,13,[31][32][33].…”

“…It is particularly interesting to compare the homozygous state of this cell line with the pESCs that are derived from artificially activated oocytes in mice [32,33] and in humans [13,34]. Artificial parthenogenesis is routinely induced by activating MII oocytes and preventing second polar body extrusion, which is thought to produce diploid embryos that are more likely to develop to the blastocyst stage [9,13,[31][32][33]. This method has been proved to produce efficient pESCs, but only predominantly heterozygous pESCs could be obtained owing to early recombination events [13,[32][33][34].…”

“…Artificial parthenogenesis is routinely induced by activating MII oocytes and preventing second polar body extrusion, which is thought to produce diploid embryos that are more likely to develop to the blastocyst stage [9,13,[31][32][33]. This method has been proved to produce efficient pESCs, but only predominantly heterozygous pESCs could be obtained owing to early recombination events [13,[32][33][34]. These cells could be used as autologous stem cells for the oocyte donor, but were of limited use for storage in the hESC bank owing to the heterozygosity of HLA loci.…”