Objective:To establish a stable and effective system of human foreskin fibroblast(HFF) feeder layer in order to culture human embryonic germ cells(EGCs) in vitro.Methods:The HFF were isolated and cultured from the 4～5 year old children’s foreskin through mechanical method and primary tissue digestion.The condition of cell growth was observed under inverted phase contrast microscope.The purity of cells

cell growth curve and cell cycle of HFF were detected by immunocytochemistry

MTT and flow cytometry.The optimal concentration of mitomycin C and the time for treating with mitomycin C on HFF were determined by MTT assay.The primordial germ cells from human embryos of 5～11 weeks were isolated;EGCs were cultured on the HFF feeder layer treated by mitomycin C and the state of cells growth was observed.Results:The mechanical method and primary tissue digestion was a credible and efficient way to obtain primary HFF

with which the cells had high purity and active proliferative abilities

and quickly entered exponential phase of growth.The cell cycle corresponded to cell growth curve.The fibroblasts

with an anticipated life span of 60 generations

were used in our experiments after 3～30 generations.HFF proliferation could be efficiently repressed after being treated with mitomycin C 12.5 mg/L for 2.5 h.Human EGCs cultured in the HFF feeder layer could form typical embryo germocyte colony and last P3 generations continuously.Conclusion:HFF feeder layer could efficiently support the growth of human EGCs.