Derivation of human embryonic stem cell lines from single blastomeres of low-quality embryos by direct plating

Abstract

To explore a simple method of establishing pluripotent human embryonic stem cell (hESC) lines from single blastomeres of low-quality (LQ) embryos. Blastomeres were isolated from normally fertilized, day-3 pre-implantation LQ embryos by dissolving of the zona pellucida and were then plated directly onto inactivated human foreskin fibroblasts. The subsequent culture was identical to that used to derive a hESC line from the inner cell mass of a blastocyst. The established hESC lines were passaged and characterized. Two hESC lines were produced by culturing the blastomeres individually in a hESC culture system (hESC-CS). Both of the hESC lines maintained a normal 46-chromosome XY karyotype, expressed stemness markers, and showed a pluripotent phenotype, including the ability to differentiate into all three germ layers in vitro and in vivo. 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 for deriving hESC lines.