体外扩增人骨髓间充质干细胞（MSCs）及其免疫抑制特性的研究

发布时间：2018-04-15 23:03

本文选题：间充质干细胞 + 混合淋巴细胞反应　；参考：《南京医科大学》2005年硕士论文

【摘要】：目的探讨体外无血清培养扩增临床规模人骨髓间充质干细胞(MSCs)及其抑制淋巴细胞增殖的免疫调节特性。方法人骨髓MSCs来自12例正常人及恶性血液病患者,采用无血清培养基体外培养。外周血淋巴细胞来自2例正常志愿者。MSCs与单向混合淋巴细胞(MLRs)和双向MLRs共培养,应用二乙酰羧基荧光素-琥珀酰亚胺酯(CFSE)标记双向MLRs中反应细胞。细胞增殖应用Cell Counting Kit-8(CCK-8)试剂盒检测,T细胞细胞亚群采用流式细胞术(FCM)分析。结果能够从所有26.3±8.8 mL骨髓样本中提取MSCs,其中10例成功扩增。扩增后MSCs表面表达CD105、CD73、CD13、CD90,而不表达骨髓造血细胞的表面抗原。MSCs呈剂量依赖性非MHC限制性抑制同种抗原诱导的淋巴细胞增殖。Th2、Tc2比例降低,Th1、Tc1比例无明显变化;T细胞早期活化标记CD69表达增强;CD4+CD25+调节性T细胞(CD4+CD25+Tr)比例增加。
[Abstract]:Objective to investigate the immunomodulatory characteristics of human bone marrow mesenchymal stem cells (MSCs) in vitro in serum-free culture.Methods Human bone marrow MSCs was cultured in serum-free medium in 12 normal subjects and patients with hematologic malignancies.Peripheral blood lymphocytes from 2 normal volunteers were co-cultured with unidirectional mixed lymphocytes (MLRs) and bidirectional MLRs. The reactive cells were labeled with diacetyl carboxyl fluorescein succinimide ester (CFSE) in bidirectional MLRs.Cell Counting Kit-8 CCK-8 kit was used to detect T cell subsets by flow cytometry (FCM).Results MSCs were extracted from all 26.3 卤8.8 mL bone marrow samples, 10 of which were successfully amplified.The surface antigen of bone marrow hematopoietic cells was not expressed on the surface of MSCs in a dose-dependent manner. MSCs inhibited the proliferation of lymphocytes induced by alloantigen in a dose-dependent manner. Th2Tc2 decreased the ratio of Th _ 1 to Tc _ 1. There was no significant change in the early survival of T cells.The expression of CD69 increased with the increase of CD4 CD25 regulatory T cell CD 4 CD25 Tr.
【学位授予单位】：南京医科大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R392

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