GROWTH OF HUMAN EMBRYONIC GERM CELLS ON HUMAN EMBRYONIC FIBROBLASTS / 解剖学报

ABSTRACT

Objective To investigate the isolation, purification and culture methods of human embryonic germ cells (EGCs) on feeder layer cells of human embryonic fibroblasts. Methods Primordial germ cells(PGCs) from the genital ridges of 6-11 weeks(post fertilization) human embryos were cultrued on feeder layer cells of human embryonic fibroblasts(HEFs) which were isolated from the 3-4 month embryos and routinely cultured for over 25 passages. The medium composed of growth factors and differentiation inhibitory factors. The cultures were analyzed with the expression of alkaline phosphatase, immunologic markers (SSEA-1,SSEA-4) and the transcription factor Oct-4 that have been used to routinely to characterize EGCS. Results A large-scale EG cells were obtained and meintained on feeder layers for over 8 passages. The cell surface marker showed that the EGCs possess high levels of AP activity. EGCs colonies stained strongly for SSEA-4,SSEA-1 and they expressed the transcription factor Oct-4.Conclusion EGCs have potential to maintain long term proliferation and undifferentiation state on human embryonic fibroblasts in vitro.;