A highly homozygous and parthenogenetic human embryonic stem cell line derived from a one-pronuclear oocyte following in vitro fertilization procedure.
Homozygous human embryonic stem cells (hESCs) are thought to be better cell sources for hESC banking because their human leukocyte antigen (HLA) haplotype would strongly increase the degree of matching for certain populations with relatively smaller cohorts of cell lines. Homozygous hESCs can be generated from parthenogenetic embryos, but only heterozygous hESCs have been established using the current strategy to artificially activate the oocyte without second polar body extrusion. Here we report the first successful derivation of a human homozygous ESC line (chHES-32) from a one-pronuclear oocyte following routine in vitro fertilization treatment. chHES-32 cells express common markers and genes with normal hESCs. They have been propagated in an undifferentiated state for more than a year (>P50) and have maintained a stable karyotype of 46, XX. When differentiated in vivo and in vitro, chHES-32 cells can form derivatives from all three embryonic germ layers. The almost undetectable expression of five paternally expressed imprinted genes and their HLA genotype identical to the oocyte donor indicated their parthenogenetic origin. Using genome-wide single-nucleotide polymorphism analysis and DNA fingerprinting, the homozygosity of chHES-32 cells was further confirmed. The results indicated that 'unwanted' one-pronuclear oocytes might be a potential source for human homozygous and parthenogenetic ESCs, and suggested an alternative strategy for obtaining homozygous hESC lines from parthenogenetic haploid oocytes.
Lin
Ge
G
Institute of Reproductive and Stem Cell Engineering, Central South University, Changsha 410078, China.
OuYang
Qi
Q
Zhou
Xiaoying
X
Gu
Yifan
Y
Yuan
Ding
D
Li
Wen
W
Liu
Gang
G
Liu
Tiancheng
T
Lu
Guangxiu
G
D000818:Animals; D002454:Cell Differentiation; D002460:Cell Line / Q000166:cytology* / Q000378:metabolism; D019070:Cell Lineage; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005307:Fertilization in Vitro; D018392:Genomic Imprinting; D006680:HLA Antigens / Q000378:metabolism; D006720:Homozygote; D006801:Humans; D007150:Immunohistochemistry; D007621:Karyotyping; D051379:Mice; D016513:Mice, SCID; D009865:Oocytes / Q000166:cytology*; D010312:Parthenogenesis; D020641:Polymorphism, Single Nucleotide
D006680:HLA Antigens