目的:探寻有效,简捷的分选出纯度较高,生物活性保存良好的外周血和蜕膜组织单核巨噬细胞,用以研究妊娠早期单核巨噬细胞功能特征及其调控因素.方法:比较运用MACS法和传统方法分离、纯化妊娠早期外周血和蜕膜组织单核巨噬细胞的效果.结果:经台盼兰染液检测细胞活性,传统贴壁法获得的单核细胞活性率为(92.50±0.57)%,MACS法获得的单核细胞活性率为(92.70±0.62)%,两者没有显著差异.传统贴壁法获得的巨噬细胞活性率为(82.40±0.64)%,MACS法获得的巨噬细胞活性率为(82.60±0.61)%,两者没有显著差异.经流式细胞仪检测细胞纯度,贴壁法获得的单核细胞纯度为(85.83±1.50)%,显著低于MACS法获得的单核细胞纯度(91.89±1.37)%.贴壁法获得的巨噬细胞纯度为(35.25±1.38)%,显著低于MACS法获得的巨噬细胞纯度(76.40±2.02)%(P<0.05).结论:MACS方法分选得到的妊娠早期外周血单核细胞和蜕膜组织巨噬细胞纯度高,生物活性保存良好.MACS方法是一种有效,简捷的分离、纯化单核巨噬细胞的方法,运用此方法,有益于更确切的描述妊娠早期参与母胎界面免疫调控的职能细胞的作用及其调节方式.
Objective: To find a simple and effective method isolating monocytes/macrophages from peripheral blood and decidua in early pregnancy. Methods: The activity and purity of monocytes/macrophages isolated with MACS and traditional method were compared. Results: By Tyrpan blue staining, The viability of monocytes isolated by traditional and MACS method was (92.50 ± 0.57)% and (92.70 ± 0.62)% respectively. No significant difference was found. The viability of macrophages isolated by traditional and MACS method was (82.40 ± 0.64)% and (82.60 ± 0.61 )% respectively. No significant difference was observed;With flow cytometry the purity of monocytes isolated by traditional method (85.83 ± 1.50)% was significantly lower than that isolated by MACS (91.89 ± 1.37)% and the purity of macrophages isolated by traditional method (35.25 ± 1.38)% was significantly lower than that isolated by MACS (76.40 ± 2.02)%, P 〈 0.05. Conclusions: MACS is a simple and effective method in isolating monocyte/ macrophages from peripheral blood and decidua in early pregnancy, which may be helpful to study the role of immune ceils in maternal-fetal immune tolerance.