骨髓间质干细胞免疫调节作用的实验研究

发布时间：2018-06-29 17:00

  本文选题：间质干细胞 + 免疫调节　； 参考：《江苏大学》2005年硕士论文

【摘要】：目的:研究骨髓间质干细胞体内外免疫调节的作用及其机制。 方法:从胚胎及Wistar大鼠股骨中分离培养间质干细胞,经克隆化分选出形态均一的细胞,用流式细胞术检测其表面标志以鉴定其纯度。将分离培养的骨髓间质干细胞接种于96孔板,待其长满孔面积的80%～90%时用直线加速器以30GY剂量照射,然后与分离的外周血淋巴细胞共培养3d,以单独培养的淋巴细胞为对照,加入ConA刺激68h,分别使用MTT法和~3H-TdR法检测MSCs对外周血淋巴细胞增殖反应的影响。采用RT-PCR和Real-time PCR检测经MSCs处理的人外周血淋巴细胞TGF-β1、IL-6、IL-8、IL-10的基因表达,以未经MSCs处理的同一人外周血淋巴细胞标本作对照。将SD大鼠的真皮移植到Wistar大鼠创口上,加压包扎,同时将2×10~6的Wistar大鼠MSCs经尾静脉注射到Wistar大鼠体内,以经同样手术处理而不注射MSCs的Wistar人鼠为对照。无菌分离Wistar大鼠外周血及脾脏淋巴细胞,并用Wistar大鼠MSCs及ConA处理,以ConA单独处理的Wistar大鼠淋巴细胞为对照。将2×10~6的MSCs在不同的时间点经尾静脉注射到Wistar大鼠体内,在每次注射前经尾静脉采血测定外周血中CD4+、CD8+淋巴细胞占整个淋巴细胞数量的比值。 结果:MSCs对ConA刺激的淋巴细胞增殖的影响与MSCs和淋巴细胞的比值有关,当比值较大时MSCs对ConA刺激的淋巴细胞增殖有明显的抑制作用,但是比值较小时反而会促进其增殖。MSCs对淋巴
[Abstract]:Objective: to study the immunomodulatory effect and mechanism of bone marrow mesenchymal stem cells (BMSCs) in vitro and in vivo. Methods: the mesenchymal stem cells were isolated from the femur of embryo and Wistar rats. The homogeneous cells were isolated by cloning. The surface markers were detected by flow cytometry to identify their purity. Bone marrow mesenchymal stem cells (BMSCs) isolated and cultured were inoculated on 96-well plate. The cells were irradiated with 30 GY with a linear accelerator for 80 ~ 90 full hole area, and then co-cultured with peripheral blood lymphocytes for 3 days. The effects of MSCs on the proliferation of peripheral blood lymphocytes were detected by MTT assay and 3H-TdR assay respectively. RT-PCR and Real-time PCR were used to detect the gene expression of TGF- 尾 1, IL-6, IL-8 and IL-10 in human peripheral blood lymphocytes treated with MSCs, and the same human peripheral blood lymphocytes without MSCs were used as controls. SD rat dermis was transplanted into the wound of Wistar rats, and the MSCs of 2 脳 10 6 Wistar rats were injected into Wistar rats via tail vein. The peripheral blood and spleen lymphocytes of Wistar rats were isolated aseptically and treated with MSCs and Cona of Wistar rats. The lymphocytes of Wistar rats treated with Cona alone were used as control. MSCs (2 脳 10 ~ (6) were injected into Wistar rats through caudal vein at different time points. Before each injection, the ratio of CD4 / CD8 lymphocytes to the total number of lymphocytes in peripheral blood was measured. Results the effect of 1: MSCs on the proliferation of Cona stimulated lymphocytes was related to the ratio of MSCs and lymphocytes. When the ratio was higher, MSCs could obviously inhibit the proliferation of Cona stimulated lymphocytes, but when the ratio was small, it would promote the proliferation of MSCs.
【学位授予单位】：江苏大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

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