系统性硬化症患者骨髓间充质干细胞的生物学特性

摘要

目的:研究系统性硬化症(SSc)患者骨髓间充质干细胞(BM-MSC)的生物学特性.方法:从骨髓中分离培养间充质干细胞(MSC),从形态、表面标记和多向分化潜能等方面进行鉴定.通过淋巴母细胞转化、混合淋巴细胞培养研究SSc患者MSC的免疫调节功能,用RT-PCR方法检测MSC免疫相关因子的表达.结果:从4例SSc患者和3例对照者骨髓中分离培养MSC,传代6次后其逐渐衰老.所分离细胞有较高纯度,90%以上处于G0/G1期.部分SSc患者表现出自发成骨倾向.细胞个数MSC:T≥1:80时,患者和对照MSC均可刺激异基因T细胞增殖(P<0.05),但当MSC:T≥1:160时抑制有丝分裂原引起T细胞增殖(P<0.05).SSc患者的MSC有免疫调节功能,通过RT-PCR证实患者MSC表达免疫相关因子白细胞介素-6(IL-6)、巨噬细胞集落刺激因子(M-CSF)、转化生长因子-β1(TGF-β1).结论:SSc患者MSC具有有限的体外扩增能力,部分患者有自发成骨倾向.SSc患者的MSC有部分免疫调节功能.%Objective To identify the characteristics of mesenchymal stem cell ( MSCs) in patients of systemic sclerosis (SSc). Methods MSCs were isolated and expanded from bone marrow samples of SSc patients and identified by morphology, phenotype and function. Mixed lymphocyte reaction and lymphocyte transformation test were performed and expression of immune cytokines was detected with RT-PCR for evaluation of immunomodulation function of MSCs. Results MSCs were successfully isolated and expanded with an aging process after 6 passages. Phenotype assay confirmed a high purity of MSCs. Cell cycle assay demonstrated that over 90% of SSc and control MSCs remained in Go/G, phase without significant difference between passages. Some patient inclined to osteogenic differentiation. Both SSc derived and control MSCs acted as stimulate to allogenic T cells when MSC: T^l: 80 (P <0. 05), but inhibited mitogen-induced T cell proliferation when MSC=T>1: 160 (P <0. 05). SSc derived MSCs presented an immune expression profile of cytokines, with reduced expression of macrophage colony-stimulating factor ( M-CSF) and transforming growth factor-pl (TGF-f$l) in some patients. Conclusion MSCs of SSc patients possessedlimited expasion and immunomodulation function.