摘要
背景:细胞替代治疗帕金森病成为目前研究的一个热点,如何进行体外诱导获得足量的有效多巴胺能神经元是治疗该疾病的一种策略。目的:探讨人脐血间充质干细胞体外诱导分化为多巴胺能神经元的可行性。方法:将传至第5代人脐血间充质干细胞以5×106L-1接种,先用20μg/L表皮生长因子+20μg/L碱性成纤维细胞生长因子预诱导24h后,分为4组:对照组加入含体积分数为10%胎牛血清的DMEM/F12培养液,不加任何诱导液;其余3组分别加入100μmol/L抗坏血酸,1μmol/L全反式维甲酸,50μg/L胶质细胞源性神经营养因子单独或联合进行诱导。结果与结论:各诱导组均表达酪氨酸羟化酶、多巴胺转运体、多巴胺受体D2 mRNA。诱导分化后的细胞能表达神经元、星形胶质细胞特异性抗原。与对照组相比,各诱导组巢蛋白、神经元特异性烯醇化酶、胶质纤维酸性蛋白、酪氨酸羟化酶、多巴胺转运体、多巴胺受体D2阳性细胞率均明显升高(P<0.05);且全反式维甲酸+胶质细胞源性神经营养因子组和3种物质联合诱导组升高幅度高于抗坏血酸组(P<0.05),3种物质联合诱导组升高幅度高于全反式维甲酸+胶质细胞源性神经营养因子组(P<0.05)。结果提示抗坏血酸,全反式维甲酸,胶质细胞源性神经营养因子单独或联合均能促进人脐血间充质干细胞向多巴胺能神经元分化,3种物质联合应用效果最高。
BACKGROUND： Parkinson＇s disease treated by cell replacement therapy has become a hot research at present, and how to obtain adequate and effective dopaminergic neurons in vitro is a strategy in the treatment of the disease. OBJECTIVE： To investigate the feasibility of human umbilical cord blood-derived mesenchymal stem cells differentiating into dopaminergic neurons in vitro. METHODS： The fifth passage human umbilical cord blood-derived mesenchymal stem cells were inoculated with 5×10^3/mL. The cells were pre-inducted with 20 ng/mL epidermal growth factor＋20 ng/mL basic fibroblast growth factor for 24 hours. Then the cells were divided into four groups： the control group was not given induction medium but only cultured in DMEM/F12 culture medium containing 10% fetal bovine serum; the other three induced groups were induced with 100 μmol/L ascorbic acid, 1 μmol/L all-trans retinoic acid 50 μg/mL glial ceil line-derived neurotrophic factor alone or in combination respectively. RESULTS AND CONCLUSION： The expression of tyrosine hydroxylase, dopamine transporter and dopamine receptor D2 mRNA could be seen in the induced groups. After induction and differentiation, the cells could express the neurons and glial cell-specific antigens. Compared with the control group, the positive rates of nestin, neuron-specific enolase, glial fibrillary acidic protein, tyrosine hydroxylase, dopamine transporter and dopamine receptor D2 in the induced groups were significantly increased （P 〈 0.05）; the positive rates in the all-trans retinoic acid＋glial cell line-derived neurotrophic factor group and the combination induced group were significantly higher than those in the ascorbic acid group （P 〈 0.05）, but the increased degree of the combination induced group was significantly higher than that of the all-trans retinoic acid＋glial cell line-derived neurotrophic factor group （P 〈 0.05）. Ascorbic acid, all-trans retinoic acid＋glial cell line-derived neurotrophic factor, all-trans retinoic acid＋glial cell line-derived neurotrophic factor＋ascorbic acid can promote the human umbilical cord blood-derived mesenchymal stern cells to differentiate into dopaminergic neurons, and the combined induction can get the best effect.
出处
《中国组织工程研究》
CAS
CSCD
2012年第49期9186-9191,共6页
Chinese Journal of Tissue Engineering Research
