AIM: In order to replace mouse embryonic fibroblast cells by human embryonic fibroblast cells to support the growth of human embryonic stem cells, the effects of human/mouse embryonic fibroblast cells on the growth of human embryonic stem cells were compared. METHODS: Both mouse and human embryonic fibroblast cells were used as feeder layer to support human embryonic stem cells. The proliferation and differentiation of human embryonic cells were observed. RESULTS: Combined use of human leukemia inhibitory factor with human/mouse feeder layer cells would support growth and proliferation of human embryonic stem cells and kept in undifferentiated condition. There was no difference between human/mouse cell. CONCLUSION: Human embryonic fibroblast cells can be used to support proliferation of human embryonic stem cells, eliminating the influence of foreign protein.