体外模拟海马神经再生微环境下的NSCs增殖和向神经元分化

目的:探讨用切割穹窿海马伞海马提取液在体外模拟体内海马神经再生微环境下,对海马神经干细胞(neural stem cells,NSCs)增殖和向神经元分化的影响。方法:分别制备正常侧和切割穹窿海马伞侧海马提取液。实验分为对照组、正常组和切割组,分别在海马NSCs中加入DMEM/F12培养基、含正常侧海马提取液及切割侧海马提取液的DMEM/F12培养基。用WST-8和Ki67/Sox2免疫荧光检测NSCs的增殖,用DCX免疫荧光检测NSCs向神经元的分化情况。结果:切割组与正常组及对照组相比,WST-8检测OD450比值明显增高(P<0.05);Ki67阳性细胞和Sox2阳性细胞的比例均明显增高(P<0.001)。结论:用切割穹窿海马伞侧海马提取液在体外模拟海马神经再生微环境,可以促进海马NSCs的增殖和向神经元的分化。

Proliferation and neuronal differentiation of NSCs by simulation of hippocampal neural regeneration in vitro

Objective: To investigate the proliferation and neuronal differentiation of NSCs,fimbria-fornix transected hippocampal extracts were used to mimic the hippocampal neural regeneration in vitro.Methods: Successfully prepared normal hippocampal extracts and transected extracts after hippocampal fimbria-fornix transection.Cells were divided into control group,normal group and transected group.DMEM/F12 medium,DMEM/F12 medium contained normal hippocampal extracts,DMEM/F12 medium contained transected hippocampal extracts were added into each group.WST-8 and Ki67/Sox2 immunofluorescence were used to detect the proliferation of NSCs,DCX immunofluorescence were used to detect neuronal differentiation.Results: In transected group,the ratio of OD450 detected by WST-8 was obviously higher than normal group and control group(P<0.05);the proportion of Ki67 and Sox2 positive cells in the transected group was obviously higher than the other groups(P<0.001).Conclusion: Using fimbria-fornix transected hippocampal extracts to simulate hippocampal neural regeneration in vitro can promote the proliferation and neurons differentiation of NSCs.