联用密度梯度离心与免疫磁珠法分离纯化人表皮朗格汉斯细胞

Separation and purification of Langerhans cells from human epidermis by density gradient centrifugation and magnetic cell sorting

目的:探讨获取较高纯度和活性的人表皮朗格汉斯细胞(LC)的实验方法。方法:联合采用密度梯度离心和免疫磁珠的方法分离纯化人表皮LC,将分选后的细胞用结合有异硫氰酸荧光素(FITC)的鼠抗人CD1a单抗标记,经流式细胞仪检测LC的纯化率,采用0.4%锥虫蓝染色检测细胞的活性。结果:分选所得LC纯化率为84.48%,锥虫蓝染色显示细胞活性>95%。结论:联用密度梯度离心和免疫磁珠法,可获得较高纯度、有活性的LC细胞,且具有操作简单、易于无菌条件下分离纯化的优点。

Objective: To improve the method of obtaining purified and viable Langerhans cells from human epidermis. Methods: Density gradient centrifugation and magnetic cell sorting assay were used simultaneously to separate and purify the Langerhans cells from human epidermis. The separated cells were labeled with FITC conjugated monoclonal mouse-anti-human CD1a antibody. The purification rate was analyzed by flow cytometry and the cell viability was detected by 0.4% trypan blue staining. Results: The purification rate of Langerhans cells was 84.48% and the cell viability was more than 95%. Conclusions: Highly purified and viable Langerhans cells could be obtained by means of density gradient centrifugation and magnetic cell sorting assay. It is a feasible approach to separate and purify Langerhans cells under sterile conditions.