Culture Of Sprague-dawley Rats' Neural Progenitor Cells In Vitro And Effects Of Egf, Bfgf, Lif

Objective:To isolate and proliferate neural progenitor cells(NPCs)from newborn Sprague-Dawley(SD)rats'hippocampus in vitro, and to observe the growth of NPCs. Then to investigate the effects of epidermal growth facto(rEGF), basic fibroblast growth factor(bFGF)and leukemia inhibitory factor(LIF)on proliferation of SD rat'NPC. Methods:The EGF/bFGF-responsive NPCs, having been isolated by mechanical method and cultivated for three passages, was divided into six groups which were supplemented with EGF, bFGF, EGF+LIF, bFGF+LIF, bFGF+EGF, bFGF+EGF+LIF respectively. The cells were identified and counted at the end of passage 9(60 d).The sum was accessed. Results:The newborn SD rats'NPCs isolated by mechanical method could be cultured and proliferated in long term in vitro. The groups supplemented with bFGF and bFGF+LIF could not be continued to culture after passage 6. The other four groups still could show the Nestin-positive neurospheres at the end of passage 9. The cells had the ability to differentiate into three neural terminal cells. The amount of NPCs in EGF+bFGF group was more than in bFGF group at the end of passage 9. The differents between bFGF group and bFGF+LIF group or between EGF+bFGF group and EGF+bFGF+LIF group were not significant(P>0.05). Conclusions:NPCs could be cultured and proliferated in long term. The group supplymented with EGF+bFGF presented stronger effect on the proliferation of SD rat's NPC than that of the group supplymented with only EGF or bFGF,and the group supplymented with bFGF presented stronger than that of the group supplymented with EGF. LIF has no effect on the proliferation of SD rat's NPC.