摘要
目的建立缺氧损伤模型,观察缺氧损伤对神经干细胞分化的影响。方法用MTT,LDH等方法比较95%N2,5%CO2比率的缺氧气体分别培养1,2,4,6,8 h神经干细胞的变化。结果三天内的大鼠海马存在大量神经干细胞,可分化为神经元、胶质细胞等神经细胞类型,95%N2,5%CO2比率的缺氧气体培养6 h后神经干细胞OD值由缺氧前0.369±0.352降至0.270±0.229,LDH+漏出量由缺氧前10.374±0.390 unit/ml.min增加到14.710±1.337 unit/ml.min,差异显著。结论证实了新生大鼠海马内提取的细胞为神经干细胞,神经干细胞缺氧培养6 h可以建立神经干细胞缺氧损伤模型,但短时间缺氧培养对细胞分化类型无明显改变。
Objective To isolate and culture NSCs from hippocampus of newly born SD rats in vitro, identify the NSCs and make hypoxia model, and investigate the regulation of oxygen deficiency on the proliferation and orientating differentiation of NSCs. Methods Immunocytochemical stainings of anti- Nestin, NF-L,GFAP were used to identify the cells. NSCs were exposed to 5% O2 for 1 h,2 hs,4 hs, 6 hs or 8 hs. Proliferation of NSCs were observed after hypoxia by MTT and LDH method and the ratio of the differentiated neurons, morphological changes were detected. Results NSCs dissociated from SD rats can be divided into neurons and astrocytes etc. After 6 hours of hypoxia,the OD value decreased from 0. 369 2± 0. 352 before anoxia to 0. 270± 0. 229, the effluxes of LDH increased from 10. 374±0. 390 unit/ml, rain before anoxia to 14. 710 ± 1. 337 unit/ml, rain. there were significant changes of the ratio of the differentiated neurons. Conclusion NSCs can be dissociated from SD rat, and After 6 hours of anoxia, hypoxia model can be made, there were no significant changes of the ratio of the differentiated neurons.
出处
《佛山科学技术学院学报（自然科学版）》
CAS
2007年第4期52-55,共4页
Journal of Foshan University(Natural Science Edition)
