免疫磁性分离技术分离、纯化小鼠表皮郎格罕细胞  

Isolation and Purification of Epidermal Langerhans Cells Using Immunomagnetic Isolation Technique

作　　者：范青源[1] 郑茂荣[1] 牟贤龙[1] 方跃明[1] 

机构地区：[1]第二军医大学附属长海医院皮肤科,上海200433

出　　处：《中华皮肤科杂志》1996年第1期49-50,共2页Chinese Journal of Dermatology

基　　金：国家自然科学基金资助课题

摘　　要：为了探讨表皮郎格罕细胞(LC)的培养方法,研究其功能特点,须将 LC 与表皮其它细胞相分离。我们在 Percoll 密度梯度离心技术纯化 LC 的基础上,采用免疫磁性分离技术,取得了更为满意的结果。通过生物素-亲合素的介导,用结合大鼠抗小鼠细胞表面 Ia 抗原的单抗的免疫磁珠吸附 LC 表面,经高强度磁场筛选,从而将 LC 从表皮细胞悬液中分离、纯化出来。结果表明,LC 的纯化率超过99%,纯化后细胞活性超过95%,产率超过80%。同种混合白细胞应答检测证实纯化后的 LC 仍有功能活性。该方法操作简单、快速,且可获得高纯化率、有功能活性的 LC。In order to investigate a method of cell culture and study the functional character of epi- dermal Langerhans cells(LC),it is necessary to separate LC from the Other epidermal cells.Based on purification of LC using Percoll density gradient centrifugation technique,we applied the immunomag- netic isolation technique to isolate and purify mouse LC,and obtained satisfactory results.By the induc- tion of avidin-biotin,we used the immunomagnetic beads directly conjugated to rat-anti-mouse Ia mono- clonal Ab to adhere to the surface of LC.The positive cells were selected from the epidermal cell sus- pension through strong magnetic field.The results showed that the enriched cell purity was up to 99%, the cell viability was up to 95% and the rate of cell recovery was up to 80%.Allogenic mixed leukocyte reaction showed that the enriched LC were functionally active.The method is technically simple and fast,and allows good recovery Of a highly purified pOpulation of LC that are functionally active.

关 键 词：郎格罕细胞 免疫磁性分离 纯化 分离 

分 类 号：R392.12[医药卫生—免疫学]