体内、体外诱导小鼠调节性树突状细胞的实验研究

发布时间：2018-03-07 13:30

本文选题：调节性树突状细胞　切入点：内皮型脾脏基质细胞　出处：《内蒙古医学院》2011年硕士论文　论文类型：学位论文

【摘要】：目的通过用体内和体外两种方法分别来诱导调节性树突状细胞的生成,同时比较了C57BL/6和NOD/LtJ两种小鼠在诱导调节性树突状细胞能力方面的差异。方法体内方法主要是通过用小鼠腹腔注射细菌,然后通过流式细胞术检测脾脏内调节性树突状细胞生成。首先观察了小鼠腹腔注射细菌数在不同的数量时对调节性树突状细胞生成的影响;其次观察了调节性树突状细胞在注射细菌后产生的不同的时间效应;同时也观察了小鼠腹腔注射LPS不同剂量时对调节性树突状细胞的诱导效应;随后又观察了小鼠腹腔分别注射细菌和改良布氏硫乙醇酸盐培养基溶液时,检测了小鼠脾脏内调节性树突状细胞生成的差异;接着比较了C57BL/6和NOD/LtJ两种小鼠注射细菌后在诱导调节性树突状细胞能力方面的差异;最后通过胞内因子染色观察了CD4~+Foxp3~+T细胞是否随调节性树突状细胞的产生而产生。体外方法是通过把连续传代的内皮型脾脏基质细胞与骨髓单个核细胞连续培养14天后,可使骨髓中的造血前体细胞分化成树突样细胞,然后通过流式细胞术对细胞表面分子进行鉴定。结果体内方法通过腹腔注射细菌可以诱导出调节性树突状细胞的产生,其中以注射细菌数为1×10~8时诱导效果最为明显;而且调节性树突状细胞的产生有一定的时间效应,其在注射细菌5天后产生的尤为显著;同时腹腔注射布氏硫乙醇酸盐培养基溶液不会诱导出调节性树突状细胞的产生;而NOD/LtJ小鼠与C57BL/6小鼠相比,更易诱导出调节性树突状细胞;只是调节性树突状细胞不会促进CD4~+Foxp3~+T细胞的产生。体外方法也同样使骨髓中的造血前体细胞分化成了调节性树突状细胞。结论通过体内和体外两种方法均可以诱导出调节性树突状细胞。
[Abstract]:Objective to induce the formation of regulatory dendritic cells in vitro and in vivo. At the same time, the difference between C57BL / 6 and NOD/LtJ mice in inducing regulatory dendritic cells was compared. Then flow cytometry was used to detect the formation of regulatory dendritic cells in the spleen. Firstly, the effect of the number of bacteria injected into the abdominal cavity on the formation of regulatory dendritic cells was observed. The time effect of regulatory dendritic cells after bacterial injection and the induction of regulatory dendritic cells by intraperitoneal injection of LPS in mice were also observed. Then we observed the difference of regulatory dendritic cells in the spleen of mice after intraperitoneal injection of bacteria and modified brucelanolide medium respectively. Then the differences in the ability of inducing regulatory dendritic cells between C57BL / 6 and NOD/LtJ mice were compared. Finally, the intracellular factor staining was used to observe whether CD4 ~ Foxp3~ T cells were produced by regulatory dendritic cells. In vitro, the endothelial splenic stromal cells and bone marrow mononuclear cells were cultured continuously for 14 days. The hematopoietic precursor cells in bone marrow were differentiated into dendritic cells and then identified by flow cytometry. Results the regulatory dendritic cells could be induced by intraperitoneal injection of bacteria in vivo. Among them, the number of injected bacteria was 1 脳 10 ~ 8:00, the induction effect was the most obvious, and the production of regulatory dendritic cells had a certain time effect, especially after 5 days of injection. At the same time, the regulatory dendritic cells were not induced by intraperitoneal injection of bruclear thioglycolate medium solution, but the regulatory dendritic cells were more easily induced in NOD/LtJ mice than in C57BL / 6 mice. Only regulatory dendritic cells did not promote the production of CD4 ~ Foxp3 ~ T cells. In vitro, the hematopoietic precursor cells in bone marrow were also differentiated into regulatory dendritic cells. Conclusion both in vivo and in vitro methods can be used to produce regulatory dendritic cells. Regulatory dendritic cells were induced.
【学位授予单位】：内蒙古医学院
【学位级别】：硕士
【学位授予年份】：2011
【分类号】：R392.1

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