本文選題：間質(zhì)干細(xì)胞 + 免疫調(diào)節(jié)　； 參考：《江蘇大學(xué)》2005年碩士論文

【摘要】：目的:研究骨髓間質(zhì)干細(xì)胞體內(nèi)外免疫調(diào)節(jié)的作用及其機(jī)制。 方法:從胚胎及Wistar大鼠股骨中分離培養(yǎng)間質(zhì)干細(xì)胞,經(jīng)克隆化分選出形態(tài)均一的細(xì)胞,用流式細(xì)胞術(shù)檢測其表面標(biāo)志以鑒定其純度。將分離培養(yǎng)的骨髓間質(zhì)干細(xì)胞接種于96孔板,待其長滿孔面積的80%～90%時(shí)用直線加速器以30GY劑量照射,然后與分離的外周血淋巴細(xì)胞共培養(yǎng)3d,以單獨(dú)培養(yǎng)的淋巴細(xì)胞為對照,加入ConA刺激68h,分別使用MTT法和~3H-TdR法檢測MSCs對外周血淋巴細(xì)胞增殖反應(yīng)的影響。采用RT-PCR和Real-time PCR檢測經(jīng)MSCs處理的人外周血淋巴細(xì)胞TGF-β1、IL-6、IL-8、IL-10的基因表達(dá),以未經(jīng)MSCs處理的同一人外周血淋巴細(xì)胞標(biāo)本作對照。將SD大鼠的真皮移植到Wistar大鼠創(chuàng)口上,加壓包扎,同時(shí)將2×10~6的Wistar大鼠MSCs經(jīng)尾靜脈注射到Wistar大鼠體內(nèi),以經(jīng)同樣手術(shù)處理而不注射MSCs的Wistar人鼠為對照。無菌分離Wistar大鼠外周血及脾臟淋巴細(xì)胞,并用Wistar大鼠MSCs及ConA處理,以ConA單獨(dú)處理的Wistar大鼠淋巴細(xì)胞為對照。將2×10~6的MSCs在不同的時(shí)間點(diǎn)經(jīng)尾靜脈注射到Wistar大鼠體內(nèi),在每次注射前經(jīng)尾靜脈采血測定外周血中CD4+、CD8+淋巴細(xì)胞占整個(gè)淋巴細(xì)胞數(shù)量的比值。 結(jié)果:MSCs對ConA刺激的淋巴細(xì)胞增殖的影響與MSCs和淋巴細(xì)胞的比值有關(guān),當(dāng)比值較大時(shí)MSCs對ConA刺激的淋巴細(xì)胞增殖有明顯的抑制作用,但是比值較小時(shí)反而會(huì)促進(jìn)其增殖。MSCs對淋巴
[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.
【學(xué)位授予單位】：江蘇大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2005
【分類號】：R392

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