We evaluated whether factors secreted by human amniotic epithelial cells (HAECs) have the neuroprotective effect on rat retinal ganglion cells (RGCs) purified by immunopanning. After culture in B27 complete medium containing B27 supplement, brain-derived neurotrophic factor (BDNF), ciliary neurotrophic factor (CNTF) and forskolin, the medium was changed to: (1). medium containing N2 supplement and forskolin (N2 basal medium); (2). medium conditioned by HAECs containing N2 supplement and forskolin (HAEC-CM); and (3). N2 basal medium containing several neurotrophic factors. HAEC-CM promoted the RGC survival compared to N2 basal medium. The effect of HAEC-CM was significantly higher than that of BDNF, neurotrophin-3 and CNTF. These results suggest that HAECs may produce unknown neuroprotective factors, suggesting its potential for the treatment of RGC degenerative diseases.