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目的:观察体外诱导新生豚鼠骨髓间充质干细胞(MSCs)分化为神经细胞过程中神经细胞Tau蛋白表达量的改变。方法:用H-DMEM培养基加胎牛血清(H-DMEM/FBS)体外培养新生豚鼠MSCs并传代扩增;分为实验组和对照组,实验组使用含细胞因子的培养基诱导MSCs向神经细胞分化,对照组加入不含细胞因子的培养基;用倒置显微镜观察2组细胞形态学改变;用ELISA法定量分析诱导分化第1、4、7、10天和第14天Tau蛋白含量的变化;免疫细胞化学法检测第14天神经元特异性烯醇化酶(NSE)和Tau蛋白的表达。结果:在细胞因子作用下豚鼠骨髓MSCs逐渐向神经细胞分化,在诱导第10天时形态上有典型神经细胞样改变;ELISA法检测发现Tau蛋白在诱导分化早期含量很低,随着诱导分化时间的延长,Tau蛋白含量逐渐升高,第10天趋于稳定,至第14天时Tau蛋白含量无明显增加;免疫细胞化学法可检测到NSE和Tau蛋白抗体阳性细胞。结论:间充质干细胞向神经细胞分化过程中Tau蛋白表达量逐渐升高,其稳定表达可能是神经细胞分化完成或成熟的标志之一。
OBJECTIVE: To observe the changes of Tau protein expression in neural cells induced by differentiation of neonatal guinea pig bone marrow mesenchymal stem cells (MSCs) into neurons in vitro. Methods: MSCs cultured in H-DMEM medium supplemented with fetal bovine serum (H-DMEM / FBS) were cultured and passaged in vitro. The MSCs were divided into experimental group and control group. MSCs were induced into neurons by using cytokine- The cells in the control group were cultured in medium containing no cytokines. The morphological changes of the two groups were observed by inverted microscope. The changes of Tau protein content in the induced differentiation day 1, 4, 7, 10 and 14 were quantified by ELISA Immunocytochemistry was used to detect the expression of neuron-specific enolase (NSE) and Tau protein on the 14th day. Results: MSCs from bone marrow of guinea pig gradually differentiated into neurons under the action of cytokines, and typical morphological changes of neurons were observed on the 10th day of induction. Tau protein was found to be low in the early stage of differentiation by ELISA. With the induction of differentiation time Tau protein content gradually increased, and tended to be stable on the 10th day. There was no significant increase of Tau protein content on the 14th day. NSE and Tau antibody positive cells could be detected by immunocytochemistry. Conclusion: The expression of Tau protein in mesenchymal stem cells differentiated into neural cells gradually increased. The stable expression of Tau may be one of the markers of the completion or maturation of neural cells.