细胞因子联合纹状体条件培养液定向诱导间充质干细胞体外分化为多巴胺能神经元的研究(英文).docVIP

细胞因子联合纹状体条件培养液定向诱导间充质干细胞体外分化为 多巴胺能神经元☆ 李 府1，马丽霞1，张乐玲1，郑立波1，陈颖杰2，吴 镇2，王世富1 Cytokines combined with striatal conditioned medium to induce differentiation of mesenchymal stem cells into dopaminergic neurons in vitro Li Fu , Ma Li-xia1, Zhang Le-ling1, Zheng Li-bo1, Chen Ying-jie2, Wu Zhen2, Wang Shi-fu1 Abstract BACKGROUND: Induction rate of bone marrow mesenchymal stem cells (BMSCs) to differentiate into dopaminergic neurons in vitro is not satisfactory. OBJECTIVE: To explore the possibility of basic fibroblast growth factor, epidermal growth factor combined with striatal conditioned medium to promote directional differentiation of MSCs into dopaminergic neurons. DESIGN, TIME AND SETTING: Cell control observation. The experiment was performed at the Hematology Laboratory of Qilu Childrens Hospital and Second Hospital of Shandong University from July 2006 to December 2007. MATERIALS: Healthy adult Wistar rats were selected for MSCs isolation, and newborn Wistar rats were selected for preparation of striatal conditioned medium. METHODS: BMSCs were harvested from healthy adult Wistar rats using attachment method for passage. Wistar rats born in 24 hours were selected, and their brain tissues were removed to prepare striatal conditioned medium. The 5th passage BMSCs were collected and pre-induced in L-DMEM containing basic fibroblast growth factor and epidermal growth factor. Twenty-four hours later, pre-induction liquor was replaced by striatal conditioned medium. MAIN OUTCOME MEASURES: Morphological changes of the stem cells were observed with the inverted phase microscope. The expressions of neuron specific enolase (NSE) and tyrosine hydroxylase (TH) were identified by immunocytochemical technique. RESULTS: The BMSC body contracted into round and spindle shape after induced by basic fibroblast growth factor and epidermal growth factor combined with striatal conditioned medium. Some neuron-like cells with prominence were seen. The immunocytochemical detection