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目的:探讨脐血单个核细胞(MNCs)体外培养前后微管相关蛋白2(MAP2)和τ蛋白mRNA的表达。方法:用密度梯度离心法分离脐血MNCs,分组培养。对照组不加任何细胞因子,细胞因子组加入EGF和bFGF,终浓度均为20μg/L,每3d半量换液1次,共14d。倒置显微镜下观察培养过程中细胞形态的变化,RT-PCR法检测脐血MNCs培养前后τ蛋白及MAP2mRNA的表达。结果:培养前,脐血MNCs胞体小呈圆形。培养后细胞因子组细胞胞体较大,有粗长突起形成,类似神经元;对照组细胞胞体较小,突起较短、较细,形似星形细胞。培养前脐血MNCsτ蛋白mRNA阴性表达,而MAP2mRNA表达阳性;培养后2者均表达阳性,且细胞因子组2者阳性表达强于对照组(P<0.05)。结论:脐血MNCs表达MAP2mRNA,经培养后同时表达τ蛋白mRNA,细胞因子能增强2者的表达。
Objective: To investigate the expression of MAP2 and Tau mRNA in cord blood mononuclear cells (MNCs) cultured in vitro. Methods: Umbilical cord blood MNCs were isolated by density gradient centrifugation and divided into groups. The control group without any cytokines, cytokines group added EGF and bFGF, the final concentration of 20μg / L, every 3d half a change, a total of 14d. The changes of cell morphology were observed under inverted microscope. The expression of tau protein and MAP2 mRNA before and after culture of cord blood MNCs were detected by RT-PCR. Results: Before culturing, the cord blood MNCs cell bodies were small and round. After culturing, the cell bodies of the cytokine group were large with protuberant protuberances and were similar to neurons. In the control group, the cell bodies of the cells were smaller and the protuberances were shorter and thinner than the astrocytes. The expression of MNCsT mRNA in cord blood before culture was negative, while the expression of MAP2 mRNA was positive. The expression of MNCsT mRNA in cord blood was positive after culture, and the expression of MNCsT mRNA in cord blood was stronger than that in control group (P <0.05). CONCLUSION: MAP2 mRNA is expressed in cord blood MNCs. After cultured, tau mRNA is also expressed. Cytokines can enhance the expression of MAP2 mRNA.