不同成熟状态小鼠树突状细胞的体外诱导

Induction of Immature or Mature Fendritic Vells from Mouse Bone Marrow in Vitro

目的:体外诱导小鼠骨髓来源的不同成熟状态树突状细胞(dendriticcells,DCs)。方法:rmGM CSF体外培养小鼠骨髓细胞,5～6d后,利用磁性细胞分离器(MACS)分离CD11c+细胞,部分细胞加入rmTNFα继续培养1～2d,通过流式细胞仪检测分别检测细胞表面标志,同种混合淋巴细胞反应检测细胞的体外刺激能力。结果:小鼠骨髓细胞经rmGM CSF培养5～6d,再经过MACS分离可获得大量未成熟DCs,细胞表面出现少量短而粗的突起,低水平表达MHCⅡ分子和共刺激分子CD80、CD86及CD40,刺激同种CD4+T细胞的混合淋巴细胞反应能力较弱;加入rmTNFα继续培养可获得成熟DCs,细胞表面出现大量细长突起,细胞表面高水平表达MHCⅡ分子和共刺激分子CD80、CD86及CD40,刺激同种CD4+T细胞的混合淋巴细胞反应能力较强。结论:通过rmGM CSF或rmGM CSF+rmTNFα体外培养体系可以获得大量未成熟DCs或成熟DCs,为进一步研究DCs的生物学功能提供实验基础。

Objective: To induce immature or mature murine dendritic cells in vitro. Methods: Bone marrow cells were cultured with rmGM-CSF, and then CD11c^+ cells were purified by MACS. The maturation state of immature DCs (imDCs) or mature DCs (mDCs) was analysis by flow cytometry (FCM) analysis and mixed lymphocyte reaction (MLR). Results: Bone marrow-derived imDCs or mDCs were successfully induced with GM-CSF in vitro. The imDCs displayed low levels of MHC class II, CD80, CD86, and CD40 moleculars, and were poor stimulators for autologous CD4^+ T cells in MLR. Conversely, the mDCs showed high levels of MHC class II, CD80, CD86, and CD40 moleculars, and were strong stimulators for autologous CD4^+ T cells in MLR. Conclusion: The large numbers of imDCs or mDCs with high purity was obtained by rmGM-CSF or rmGM-CSF and rmTNFα. It will be useful for future biological studies of DCs.