中文摘要：

目的体外培养脑白质胶质源性神经祖细胞并制备神经祖细胞的缺血模型，以期进一步研究脑白质缺血性损伤时脑白质的再生机制。方法分离3日龄内SD新生大鼠的双侧脑室周围白质组织，原代培养脑白质神经祖细胞，每3-4天传代一次，并对神经祖细胞进行诱导分化培养。免疫细胞化学法分别鉴定原代和传代培养的神经球标志物神经胶质抗原2（NG21和诱导分化后少突胶质细胞前体标志物04的表达。应用自行研制的细胞缺氧箱，对传至第3-4代的神经祖细胞制备氧糖剥夺缺血性模型。建模后24h应用Hoechst33342／PI双染和CCK-8法分别评估氧糖剥夺30min、45min、60min、2h及3h神经祖细胞存活率的变化。结果脑白质神经祖细胞具有形成神经球和连续传代的能力，原代和传代培养的神经球均显示NG2阳性，并可被诱导分化为04阳性的少突胶质细胞前体。正常培养的神经祖细胞无凋亡和坏死细胞，随着氧糖剥夺时间的逐步延长，凋亡细胞逐渐增多并出现坏死细胞。不同氧糖剥夺时间组细胞存活率均不同．而且随着氧糖剥夺时间的延长，细胞存活率下降，差异有统计学意义（P〈0．05）。结论成功培养了脑白质胶质源性神经祖细胞并建立了有效可靠的祖细胞氧糖剥夺模型，为进一步研究脑白质缺血性损伤时脑白质的神经再生机制奠定了基础。

英文摘要：

Objective To in vitro culture glial lineage progenitor cells derived from periventricular white matter in neonatal rats, and establish oxygen-glucose deprivation models of progenitor cells to further explore the mechanism of neurogenesis in the ischemic white matter injury. Methods The progenitor cells were isolated and cultured from the white matter of 3-d-old neonatal rats. Cells were passaged every 3 or 4 d, and were further induced differentiation. The primary and passaged neurospheres and their differentiated cells were respectively identified by immunocytochemical stanning of NG2 （Neurospheres marker） and 04 （oligodendrocyte precursor marker）. The third- or fourth-generation progenitor cells were used to establish the ischemic model of oxygen-glucose deprivation （OGD）. The influence of different OGD periods （30, 45 and 60 min, 2 and 3 h） in the survival rates of progenitor cells was assessed 24 h after success of model making by both CCK-8 and Hoechst 33342/PI stainning. Results The progenitor cells obtained from the white matter had the capacity of forming neurospheres and reproduction which showed NG2 immunoreactive positivity, and could be induced differentiation into O4-positive oligodendrocyte precursors. No apoptotic or necrotic cells were observed in normal cultured NG2-positive progenitor cells. The apoptotic and necrotic cells tended to increase significantly with the extension of OGD period gradually. The survival rates of progenitor Cellsin turn were （85.94±3.06） % for 30 min OGD, （62.68±2.66） % for 45 rain OGD, （45.09±2.24） % for 60 min OGD, （36.70±2.84） % for 2 h OGD and （22.01±3.00） % for 3 h OGD, respectively, with significant differences between each two groups （P〈0.05）. Conclusion The oxygen-glucose deprivation models of glial lineage progenitor ceils derived from periventricular white matter of neonatal rats are successfully established, which may provide the foundation for further exploring the mechanism of neurogenesis in the ischemic white