缺氧对体外培养的鼠胚神经干细胞分化的影响

目的分离大鼠胚胎皮质神经干细胞（NSCs）进行体外培养,探讨缺氧对体外培养的鼠胚大脑皮质神经干细胞分化的影响。方法对孕14d（E14d）大鼠取鼠胚脑皮质用无血清培养技术分离培养神经干细胞，血清：培养基贴壁诱导分化。通过免疫细胞化学染色检测细胞分化情况；NSCs采用三气培养箱予以不同缺氧干预，缺氧培养后再用10％胎牛血清培养基进行贴壁分化，用MAP2免疫荧光染色检测缺氧对NSCs向神经元方向分化的影响。结果①大鼠胚胎脊髓可成功分离神经干细胞，分化后可表达神经元、星形胶质细胞的特异性抗原；②缺氧干预实验中，5％O2组尤以72h组可诱导NSCs向神经元方向分化。结论从大鼠胚胎皮质可成功分离NSCs，缺氧可影响NSCs的分化，适度缺氧可诱导离体培养的大鼠脑皮质NSCs向神经元方向分化。

The Effects of Hypoxia on the Differentiation of Cultured Nscs from Embryonic Rats in Vitro

Objective To isolate and culture neural stem cells （NSCs） from rat embryonic brain cortex and explore the effects of hypoxia on the differentiation of the NSCs in vitro. Methods The NSCs from brain cortex of E14d SD rats were isolated. The NSCs were cultured in vitro in the serum - free medium at first, and then differentiation with 10% FBS medium. The P3 NSCs were divided into experimental groups（ 10% O2 ,5% O2） and normal control group. The cells in experimental groups were cultured with serum - free medium in incubator for 7 days first in 5% O2 and 10% O2 for 24,72 or 120h,then in normal O2. Then NSCs of different groups were differentiated cultured with 10% FBS medium. The NSCs activity was detected by light microscope,and the nestin and neural cell specific antigens were detected using immunocytochemistry or immumofluorence staining. Results 5% O2 groups, especially 72h group could promote NSCs to differentiate into neurons. Conclusion Our results indicate that hypoxia may have great effect on the natural differentiation of NSCs. In vitro,fitness hypoxia may promote NSCs to differentiate into neurons.