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本研究旨在通过培养并观察C57BL/6J小鼠骨髓来源树突状细胞(BMDCs)集落、形态、表型的动态变化,为BMDCs的形态、表型等方面提供参考数据。取C57BL/6J小鼠骨髓,用粒细胞-巨噬细胞集落刺激因子(GM-CSF)诱导和培养BMDCs,在第3、6、9、12天拍照、HE染色观察BMDCs的集落、形态;FACS分析第3、6、9、12天以及LPS刺激后BMDCs的CD11c、CD80和CD86的表达;扫面电镜拍照LPS刺激后BMDCs的树突与形态。发现GM-CSF诱导和培养的C57BL/6J小鼠BMDCs在第3天形成明显的集落、第6天集落释放较多的BMDCs;显微镜下HE染色显示,第3天可见BMDCs有突起、第6天明显,第12天有明显突起的细胞数量增多;FACS分析显示,随着培养时间的增加,BMDCs的CD11c、CD80、CD86的表达逐渐上调,第12天表达较明显,LPS刺激能提高CD80、CD86的表达;扫描电镜显示,LPS刺激的BMDCs有明显的树突状突起。因此GM-CSF诱导和培养的C57BL/6J小鼠BMDC具有典型的树突状突起、表达CD11c、CD80、CD86等树突状细胞(DCs)分子标记,LPS刺激能促进CD11c、CD80、CD86的表达。
This study aimed to provide reference data for the morphological and phenotypic characteristics of BMDCs by culturing and observing the dynamic changes of the colony, morphology and phenotype of bone marrow-derived dendritic cells (BMDCs) in C57BL / 6J mice. Bone marrow of C57BL / 6J mice was used to induce and culture BMDCs with granulocyte-macrophage colony stimulating factor (GM-CSF). The BMDCs were photographed on the 3rd, 6th, 9th and 12th days. The morphology and morphology of BMDCs were observed by HE staining. The expression of CD11c, CD80 and CD86 in BMDCs after 3, 6, 9, 12 days and LPS stimulation were analyzed. The dendrites and morphology of BMDCs after LPS stimulation were examined by scanning electron microscopy. It was found that BMDCs of C57BL / 6J mice induced and cultured by GM-CSF formed obvious colonies on the 3rd day and released more BMDCs on the 6th day. HE staining on the 6th day showed that BMDCs protruded on the 3rd day, Obviously, the number of cells with obvious protrusions increased on the 12th day. FACS analysis showed that the expression of CD11c, CD80 and CD86 of BMDCs gradually increased with the increase of culture time, and the expression of CD11c, CD80 and CD86 was more obvious on the 12th day. The results of scanning electron microscopy showed that dendritic protrusions were observed in LPS-stimulated BMDCs. Therefore, BMDC of C57BL / 6J mice induced and cultured by GM-CSF has typical dendritic protuberances and expressed dendritic cells (DCs) markers such as CD11c, CD80 and CD86. LPS stimulation can promote the expression of CD11c, CD80 and CD86 .