Purpose To explore a simple method of establishing pluripotent human embryonic stem cell (hESC) lines from single blastomeres of low-quality (LQ) embryos.Methods Blastomeres were isolated from normally fertilized day 3 pre-implantation LQ embryos by zona pellucida dissolving, and were then plated directly onto inactivated human foreskin fibroblasts.The subsequent culture was identical to that of derivation of a hESC line from the inner cell mass of a blastocyst.The established hESC lines were passaged and characterized.Results Two hESC lines werc produced without cavitations by culturing the blastomeres individually in a hESC culture system(hESC-CS).Both of the hESC lines maintained a normal 46, XY karyotype, expressed stemness markers and showed a pluripotent phenotype including the ability to differentiate into all three germ layers in vitro.Conclusion The blastomeres of LQ embryos have a developmental capacity that necessitates prolonged culture.Plating of blastomeres from LQ embryos directly into the hESC-CS is a feasible method to derive hESC lines.