常氧与低氧环境下白细胞介素-1β体外诱导鼠胚中脑神经干细胞向多巴胺能神经元的分化  

In vitro differentiation of mouse embryonic mesencephalon derived neural stem cells into dopaminergic neuron induced by interleukin 1β in normoxia and hypoxia

作　　者：罗特坚[1] 丁继固[2] 

机构地区：[1]邵阳医学高等专科学校解剖学教研室,邵阳422000 [2]湖北科技学院人体解剖学教研室,咸宁437100

出　　处：《解剖学杂志》2013年第1期56-60,共5页Chinese Journal of Anatomy

基　　金：湖北省教育厅重点课题（D20092802）;

摘　　要：目的：探讨白细胞介素-1＆beta;（IL-1＆beta;）在低氧环境下体外诱导中脑源性神经干细胞的分化。方法：分离孕12d胎鼠中脑组织，制成单细胞悬液，先在无血清培养条件下做原代和传代培养；对原代和传代培养分化细胞分别行免疫细胞化学显色鉴定。鉴定后，传代培养的中脑源性神经干细胞按组接种于含10% FBS的DMEM/F12培养基和含10%FBS的DMEM/F12＋IL-1＆beta;培养基；分别置于常氧（21% O2）和低氧（3% O2）环境下诱导分化，诱导9～11 d后行小鼠抗大鼠TH免疫细胞化学显色，并行流式细胞术检测酪氨酸羟化酶（TH）及烯醇化酶（NSE）阳性细胞率。结果：各实验组中，常氧组（对照组）、常氧＋IL-1＆beta;组、低氧组、低氧＋IL-1＆beta;组TH阳性神经元分别为3.63＆plusmn;0.76、 11.76＆plusmn;0.54、 15.83＆plusmn;1.15、 23.87＆plusmn;1.83；NSE阳性神经元分别为19.82＆plusmn;0.65、 29.33＆plusmn;0.84、 41.62＆plusmn;1.51和60.35＆plusmn;1.56。各组与对照组比较，差异均有统计学意义。结论：孕12d胚鼠腹侧中脑组织可以在体外培养传代、并能诱导分化成多巴胺能神经元；在低氧环境或低氧＋IL-1＆beta;诱导下，分化率均高于常氧组，其表型更成熟。说明在低氧或低氧＋IL-1＆beta;诱导下，可明显促进中脑源性干细胞分化为形态及功能成熟的多巴胺能神经元。Objective： To explore the in vitro differentiation of neural stem cells derived from mice embryonic midbrain. Meth- ods： The midbrain tissue harvested from mice fetus at embryonic day 12 was prepared to single cell suspension, and then cul- tured and subcuhured in serum-free media. And, the cells were characterized immunocytochemically. Subsequently, the mes- encephalon derived neural stem cells were incubated in the DMEM/F12 media containing 10% fetal bovine serum （FBS） with or without interleukin 1β （IL1β）, respectively. Afterwards, these ceils were replaced into incubator containing 21% oxygen （normoxia） and 3% oxygen （hypoxia）. Nine to eleven days later, the induced cells were visualized immunocytochemically against tyrosine hydroxylase （TH）, and percentages of positive TH and NSE cells were identified by flow cytometry. Results： In normoxia only （control）, normoxia＋IL1β, hyoXia only, hypoxia＋IL1β groups, the percentage of positive TH cells were 3.63±0. 76, 11.76±0.54, 15.83±1.15 and 23.87±1.83, while the percentage of positive NSE cells were 19. 82±0. 65, 29. 33±0. 84, 41. 62±1.51 and 60. 35± 1.56, respectively. Statistical differences were detectable when percentage in other groups compared with those in control. Conclusion： The midbrain tissue from mice fetus at embryonic day 12 can be cultured and suhcultured in vitro, and differentiated into the dopaminergie neuron. Higher differentiation rated in hypoxia with and without IL1β suggests that hypoxia with and without IL1β may improve the differentiation of mice embryonic mesencephalon derived neural stem cells into dopaminergic neuron.

关 键 词：低氧 白细胞介素-1Β 中脑源性干细胞 酪氨酸羟化酶 多巴胺能神经元 

分 类 号：R392.12[医药卫生—免疫学]