Human parthenogenetic embryonic stem cells (pESCs) were previously thought to be better cell sources for ESC banking because the homozygous Human Leukocyte Antigen (HLA) would strongly increase the matching degree for certain population with relatively smaller cohorts of cell lines. While recent studies have shown that upon currently used strategy to artificial activate oocyte without second polar body extrusion, most pESCs derived would be predominantly heterozygous due to early recombination events in oocyte. Here we reported successful derivation of human homozygous and heterozygous pESC lines from one pronuclear zygote following routine in vitro fertilization treatment and artificial parthenoactivation. The hESC characters, SNPs genotyping, gene expression profiles were conducted and compared to other normal hESCs. The in vitro differentiation ability and the epigenetic stability of imprinted genes of these two cell lines before and post differentiation were further evaluated.