Human embryonic germ cells isolation from early stages of post-implantation embryos

Human embryonic germ (hEG) cell is a very important alternative pluripotent stem cell resource. We describe the derivation of hEG cells from human embryonic fetal gonads over 6-8 weeks postconception. A large number of EG-like cell clumps were obtained at passage 1 and thus facilitated the following routine culture when the donor tissues were trypsinized with gentle pipetting and plated on feeder layer cells in the initial culture. Eight diploid hEG cell lines have been cultivated in vitro for extended periods while maintaining expression of markers characteristic of pluripotent stem cells. Human EG cells expressed transcription factor Oct4, a marker of pluripotency in mouse EG cells, at a high and steady level. Expression of markers indicative of differentiation along the three germ lineages was also observed in EBs. High level of alkaline phosphatase activity was shown in EG cells. These encouraging findings provide a starting point for potential applicability of hEG cells.