【摘要】 目的探讨星形胶质细胞能否在体外诱导成年大鼠骨髓基质细胞向神经元方向分化。方法采用新生鼠海马组织来源的星形胶质细胞与转染有绿色荧光蛋白基因的骨髓基质细胞(MSCs)进行共培养,并分成三组:共培养组、共培养+脑源性神经营养因子(BDNF)组、单纯碱性成纤维生长因子(bFGF)+BDNF诱导分化组,在相差显微镜下每日观察、记录MSCs的诱导分化状况,并应用免疫荧光染色技术对分化后的MSCs进行鉴定,同时应用流式细胞术测定MSCs分化前后及星形胶质细胞的DNA含量。结果共培养第4天,部分MSCs已初步具备神经元形态:折光性强的锥形或圆形胞体及长的多极突起,免疫荧光染色呈微管相关蛋白(MAP-2ab)、神经元特异性烯醇化酶(NSE)阳性;DNA的含量测定结果显示诱导分化后第7天未发现有四倍体、六倍体细胞。结论(1)星形胶质细胞可以在体外诱导成年大鼠MSCs向神经元方向分化,这种分化并不是由细胞融合引起的。(2)星形胶质细胞具有调控神经元的分化、促进神经元成熟的功能。更多还原

【Abstract】 Objective To study whether adult marrow stromal cells can be induced to differentiate into neurons by astrocytes. Methods Adult rat MSCs infected with a retrovirus engineered to express green fluorescent protein (GFP) were co-cultured with astrocytes isolated from the hippocampus of newborn rat . Three groups were divided into: co-culture, co-culture and BDNF, control group. Then phase-contrast microscope was used to observe the differentiation of MSCs every day and the technique of immunocytofluorescence was applied to identify the differentiated MSCs. Meanwhile, Flow Cytomytry was performed to measure the DNA content of differentiated MSCs, normal MSCs and astrocytes. Results On the forth day of co-culture, some MSCs had already the typical morphological neuronal traits: the refractile and cone-like or spherical cell bodies and long processes , expressed neuron-specific enolase(NSE)and Map-2ab, a marker of mature neurons. No tetraploid or hexaploid cell was found by flow cytomytry. Conclusions (1)Mature astrocytes may induce MSCs to differentiate into neurons in vitro, which wasn’t resulted from cell fusion.(2)Astrocyte can regulate the differentiation and promote the maturation of neurons .更多还原