本文選題：間充質(zhì)干細(xì)胞 + 骨髓　； 參考：《山東大學(xué)》2006年碩士論文

【摘要】：目的：研究骨髓間充質(zhì)干細(xì)胞(Mesenchymal Stem Cells，MSCs)對(duì)PHA或異體抗原作用下的T淋巴細(xì)胞分泌功能的免疫調(diào)節(jié)作用，初步探討MSCs誘導(dǎo)免疫耐受，降低移植物抗宿主病(graft-versus-host disease，GVHD)發(fā)生率的機(jī)制。 研究方法：通過(guò)Ficoll分離法分離骨髓單個(gè)核細(xì)胞，體外培養(yǎng)擴(kuò)增出MSCs，并通過(guò)形態(tài)學(xué)特征及流式細(xì)胞術(shù)檢測(cè)其表面標(biāo)志加以鑒定。通過(guò)Ficoll分離法和尼龍棉柱法獲取外周血T淋巴細(xì)胞，將不同數(shù)量的MSCs與PHA激活的T細(xì)胞共培養(yǎng)4天，，收集培養(yǎng)上清液；將不同數(shù)量的MSCs加入混合淋巴細(xì)胞反應(yīng)(mixed lymphocyte reaction，MLR)體系共培養(yǎng)7天，收集培養(yǎng)上清液；將不同濃度的MSCs培養(yǎng)上清加入MLR體系共培養(yǎng)7天，收集培養(yǎng)上清液。應(yīng)用酶聯(lián)免疫吸附試驗(yàn)(enzyme-linked immunosorbent assay，ELISA)分別檢測(cè)各培養(yǎng)上清液中T細(xì)胞分泌的細(xì)胞因子水平，包括IL-2和INF-γ(主要由Th1細(xì)胞分泌)，IL-4和IL-10(主要由Th2細(xì)胞分泌)。 結(jié)果：骨髓單個(gè)核細(xì)胞接種于培養(yǎng)瓶后，2～4天可見成纖維樣貼壁細(xì)胞逐漸出現(xiàn)，增殖速度較慢；7～11天散在的成纖維樣貼壁細(xì)胞增殖較快，細(xì)胞形態(tài)均一；2周左右，細(xì)胞生長(zhǎng)呈漩渦狀或平行狀排列，細(xì)胞融合可到達(dá)90％。傳代細(xì)胞生長(zhǎng)迅速，主要呈成纖維樣，約第7～10天可達(dá)到融合。流式細(xì)胞儀檢測(cè)人骨髓MSCs表面抗原高表達(dá)CD29、CD105、CD166；而低表達(dá)CD34、CD45、HLA-DR。
[Abstract]:Objective: To study the immunoregulation effect of Mesenchymal Stem Cells (MSCs) on the secretory function of T lymphocytes under the action of PHA or allogenic antigen, and to explore the mechanism of MSCs induction of immune tolerance and the reduction of the incidence of graft versus host disease (graft-versus-host disease, GVHD).
Methods: Ficoll separation method was used to separate bone marrow mononuclear cells. MSCs was cultured and amplified in vitro. The morphological characteristics and flow cytometry were used to identify the surface markers. T lymphocytes were obtained by Ficoll separation and nylon cotton column method, and the number of MSCs and PHA activated T cells were collected for 4 days. The supernatant was cultured, and different amounts of MSCs were added to the mixed lymphocyte reaction (mixed
Lymphocyte reaction, MLR) system was cultured for 7 days, and the supernatant was collected and cultured. The culture supernatant of different concentration of MSCs was added to MLR system for 7 days to collect culture supernatant. The level of cytokines secreted by T cells in each culture supernatant was detected by enzyme linked immunosorbent assay (enzyme-linked immunosorbent assay, ELISA), including I. L-2 and INF- gamma (mainly secreted by Th1 cells), IL-4 and IL-10 (mainly secreted by Th2 cells).
Results: after the bone marrow mononuclear cells were inoculated in the culture bottle, the fibroid adherent cells appeared gradually and the proliferation rate was slow on the 2~4 day, and the proliferation of fibroid adherent cells on the 7~11 day was faster and the cell morphology was uniform, and the cell growth was whirlpool or parallel in 2 weeks, and the cell fusion could reach the growth of the 90%. passages cells. Rapid, mainly fibrous like, about seventh to 10 days to achieve fusion. Flow cytometry detection of human bone marrow MSCs surface antigen high expression of CD29, CD105, CD166, and low expression of CD34, CD45, HLA-DR.

【學(xué)位授予單位】：山東大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R329.2

【參考文獻(xiàn)】

相關(guān)期刊論文 前10條

1 江小霞,張毅,李秀森,吳英,于曉Y

本文編號(hào)：1864717