羧基荧光素乙酰乙酸与四甲基偶氮唑盐法检测人脐血间充质干细胞刺激脾淋巴细胞增殖作用的比较

目的 比较羧基荧光素乙酰乙酸(CFSE)与四甲基偶氮唑盐(MTT)比色法检测人脐血间充质干细胞(CB-MSCs)和神经分化的CB-MSCs对大鼠脾淋巴细胞(LCs)刺激作用的敏感程度. 方法 分别制备刺激细胞和LCs,将刺激细胞分为4组:1. CB-MSCs组;2. Dif-CB-HSCs组:脐血间充质干细胞培养7d后维甲酸(RA)处理4d诱导其神经分化细胞;3. SH-SY5Y组:人源SH-SY5Y细胞(人神经母细胞瘤细胞系)作为阳性对照组;4. Auto-LC组:自体大鼠LCs作为阴性对照组.分别将上述各组刺激细胞与LCs进行混合培养,分别通过酶标仪(MTT法),流式细胞术(CFSE法)检测LCs增殖情况( n =3). 结果 CFSE法和MTT法检测结果 均显示,CB-MSCs组和Dif-CB-MSCs组刺激LCs增殖明显弱于阳性对照组(SH-SY5Y细胞组),但CFSE法检测结果 显示,CB-MSCs组和Dif-CB-MSCs组细胞刺激LCs增殖百分率高于阴性对照Auto-LC组,MTT法检测结果 显示,CB-MSCs组和Dif-CB-MSCs组的吸光度( A )值稍低于阴性对照Auto-LC组. 结论 CFSE法比MTT法能更加客观地反映淋巴细胞增殖情况,在实际应用中更具有优势.

Compararative investigation of proliferation of lymphocytes stimulated by human umbilical cord blood mesenchymal stem cells with CFSE and MTT

Objective To compare the sensitivities of the stimulation effect of human cord blood mesenchymal stem cells(CB-MSCs) and CB-MSCs of neuronal differentiation to lymphocytes(LCs) detected with carboxyfluorescein didcetate(CFSE) and MTT. Methods To prepare LCs from SD rat and divided into four group stimulating cells: 1. CB-MSCs;2. Dif-CB-HSCs;3. SH-SY5Y(positive control);4. Auto-LC(negative control).Stimulating cells were respectively Co-cultured with LCs. The proliferation of LCs was detected with MTT and CFSE ( n =3). Results CB-MSCs and Dif-CB-HSCs stimulated LCs to proliferate more weakly than positive control detected with MTT and CFSE. The quantity of proliferation of lymphocytes Co-cultured with CB-MSCs and Dif-CB-HSCs were higher than that of Auto-LC detected with CFSE. But MTT OD value of CB-MSCs and Dif-CB-HSCs was a little lower than that of Auto-LC. Statistical analysis results showed no significant difference.Conclusion CFSE can reflect proliferation status of lymphocytes better than MTT. CFSE shows more advantages in practical use.