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目的: 体外诱导和扩增大鼠骨髓树突状细胞 (DC)的方法, 并进行生物学特性鉴定。方法: 将大鼠骨髓细胞培养 48h后, 去除悬浮细胞, 加入IL- 4和GM- CSF培养 2wk。在光镜和透射电镜下观察培养的DC的形态学特征。应用流式细胞仪检测DC表面分子MHC-Ⅱ类分子、B7 -1、B7- 2的表达水平。将DC与同种异体T细胞混合培养, 采用MTT比色法测定不同细胞密度的DC刺激同种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DC的典型形态。培养 2wk的DC表面MHC -Ⅱ类分子表达的阳性率为 74. 2%, B7- 1、B7 -2分别为 81%、76%。培养的DC具有明显刺激同种异体T细胞增殖的能力。结论: 将大鼠骨髓细胞经贴壁去除悬浮的细胞, 加入IL -4和GM -CSF培养, 可获得足量、较高纯度的DC, 为研究DC的功能奠定了基础。
OBJECTIVE: To induce and amplify rat bone marrow dendritic cells (DCs) in vitro and to identify their biological characteristics. Methods: After cultured rat bone marrow cells for 48 h, suspended cells were removed and cultured for 2 weeks with addition of IL-4 and GM-CSF. Morphological characteristics of cultured DCs were observed under light and transmission electron microscopy. Flow cytometry was used to detect the expression of MHC class Ⅱ molecules, B7 -1 and B7-2 on DCs. DC and allogeneic T cells were mixed and cultured. The ability of DCs with different cell densities to stimulate the proliferation of allogeneic T cells was measured by MTT colorimetry. Results: The cultured DC cytoplasm had large, long, dendritic shape with typical morphology of DC. The positive rate of expression of MHC-II molecules on DCs cultured for 2 weeks was 74.2%, while that of B7-1 and B7-2 was 81% and 76% respectively. DCs cultured have the ability to significantly stimulate the proliferation of allogeneic T cells. CONCLUSION: The suspension of rat bone marrow cells adherent cells, adding IL-4 and GM-CSF culture, can obtain sufficient and high purity of DC, which laid the foundation for the study of DC function.