Abstract: Objective:To observe the Results of isolation and culture of retinal stem cell in the neonatal rat and investigate their biological characteristics． Methods:The mechanical enzyme separation in vitro amplification of rat retinal stem cells was done，the capacity of cell proliferation and growth curve of CCK-8 detection were observed;cell differentiation was induced，then the expressions of nestin，bromodeoxyuridine，microtubule-associated protein-2，glial fibrillary acidic protein and thymus glycoprotein-1． 1 in cells were detected by immunocytochemical method． Results:The primary retinal stem cells formed a large number of cells in the ball suspended growth，the passaged cells could also form a ball of cells，and behaved as linear growth，the nestin and bromodeoxyuridine expressions were positive;glial fibrillary acidic protein，microtubule-associated protein-2，thymus glycoprotein-1． 1 were positively expressed in part of cells after induction of differentiation． Conclusions:The neonatal SD rat retinal stem cells can be isolated，cultured，and amplified in vitro，expressed the characteristics of stem cells，it can differentiate into retinal ganglion cells．