Percoll密度梯度离心技术分离纯化豚鼠表皮Langerhans细胞　

Isolaton and Purification of Epidermal Langerhans Cells from Guinea--pig with Percoll Density Gradient Centrifugation Technique

表皮Ｌａｎｇｅｒｈａｎｓ细胞（ＬＣ）仅占表皮细胞的２％～４％，但在免疫系统中占重要作用。为了对ＬＣ的表型、功能特点及分子生物学水平等作进一步研究，有必要获得高纯化率且具有活性的ＬＣ。我们采用连续、间断Ｐｅｒｃｏｌｌ密度梯度离心相结合的方法分离、纯化豚鼠表皮ＬＣ。结果表明ＬＣ纯化率超过６０％，产率超过８０％，细胞活性超过９０％。自体混合白细胞应答检测证实纯化的ＬＣ仍保持功能活性。该方法简单、快速、价廉，且获得非表面标记性的ＬＣ。

Although epidermal Langerhans cells (LC) account for only 2%~4% of epidermal cells,they play an important role in the immune system. In order to investigate the phenotype,functional feature, and molecular biology of LC,it is necessary to obtain the LCs with high purity and activity. A combination of continuous and interval Percoll density gradient centrifugation technique was used to isolate and purify epidermal LC from guinea-pig. The results showed that the LC purity rate was more than 60%,yield rate more than 80 %,and cell activity more than 90 %. Auto--mixed leucocyte response assay showed that the functional activity of LC remained. It is the author's opinion that this method is not only simple, quick,and cheap,but can obtain LC with no surface marker.