中华皮肤科杂志 ›› 1996, Vol. 29 ›› Issue (1): 48-50.

• 论著 • 上一篇    下一篇

免疫磁性分离技术分离、纯化小鼠表皮郎格罕细胞

范青源, 郑茂荣, 牟贤龙, 方跃明   

  1. 第二军医大学附属长海医院皮肤科, 上海200433
  • 收稿日期:1995-01-23 修回日期:1995-08-04 出版日期:1996-02-15 发布日期:1996-02-15
  • 基金资助:
    国家自然科学基金资助课题

Isolation and Purification of Epidermal Langerhans Cells Using Immunomagnetic Isolation Technique

Fan Qingyuan, Zheng Maorong, Mu Xianlong   

  1. Changhai Hospital, Second Military Medical University, Shanghai 200433
  • Received:1995-01-23 Revised:1995-08-04 Online:1996-02-15 Published:1996-02-15

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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。

关键词: 郎格罕细胞, 免疫磁性分离

Abstract: In order to investigate a method of cell culture and study the functional character of epidermal 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 induction of avidin-biotin,we used the immunomagnetic beads directly conjugated to rat-antimouse Ia mono-clonal Ab to adhere to the surface of LC.The positive cells were selected from the epidermal cell suspension 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.

Key words: Langerhans cells, Immunomagnetic isolation