大鼠树突状细胞的体外扩增与初步鉴定

目的建立体外诱导和扩增大鼠骨髓树突状细胞(DCs)的方法，并进行生物学鉴定。方法大鼠骨髓细胞经贴壁去除悬浮细胞，加入细胞因子(IL-4和GM—CSF)培养2周。经过先镜，透射电镜观察培养DCs的形态学特征；通过同种T细胞混合培养，采用MTT比色分析法测定不同浓度的DCs发同种T细胞增殖的能力。结果培养的DC胞浆突起大而长，呈树突状，具有DCs的典型形态，并且具有强烈的激发同种异体T细胞增殖的能力。结论采用大鼠骨髓细胞经贴壁去除悬浮细胞，加入细胞因子(IL-4和GM—CSF)序贯培养，能获得大量、较高纯度的DCs。

Amplication and identification of dendritic cells from rat bone marrow in vitro

To establish the method of inducing and proliferating dentritic cells (DCs) from rat bone marrow in vitro and analyze its biological characteristics. Bone marrow cells were flushed from femurs and tibias of healthy Lewis rats, depleted of erythrocytes with osmotic lysis. Cultured for 48 hours, the floating cells were removed and the adherent cells were continued to culture with cytokines including IL-4 and GM-CSF. After 14 days culture, the induced DCs were observed by photics microscope and electron microscope, also co-cultured with native T cells derived from spleen, its stimulating function was detected by MTT assay. The induced DCs had the typical morphology characteristitics. Only a little number of the induced DCs can stimulate the proliferation of allogeneic T cells efficiently and they were potent stimulators in allogeneic mixed lymphocyte reaction (MLR). Conclusion] It is an applicable method by using rat bone marrow as a source after adherent and being cultured in the medium containing IL-4 and GM-CSF, which could generate plenty of DCs with high purity and biological functions.