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骨髓和脂肪来源的人骨髓间充质干细胞在早期和晚期传代中分泌的免疫调节细胞因子的基因表达谱。

Gene expression profile of immunoregulatory cytokines secreted from bone marrow and adipose derived human mesenchymal stem cells in early and late passages.

机构信息

Basic and Molecular Epidemiology of Gastrointestinal Disorders Research Center, Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

Department of Molecular and Cell Biology, Faculty of Life Sciences and Biotechnology, Shahid Beheshti University, Tehran, Iran.

出版信息

Mol Biol Rep. 2020 Mar;47(3):1723-1732. doi: 10.1007/s11033-020-05264-2. Epub 2020 Feb 10.

Abstract

Mesenchymal Stem Cells (MSCs) have therapeutic potential in a variety of diseases; however, the safety and efficacy of their use remain ambiguous. Clinical applications of MSCs are under intensive investigation due to their immunomodulatory features and lack of immune reactivity. Therefore, having a clear perspective on the exact mechanisms underlying the regulation of cytokine secretion in different microenvironments seems crucially important.In the current study, samples from human bone marrow and adipose were collected, and peripheral blood mononuclear cells (PBMCs) were isolated and cultured in conventional medium. After MSC expansion, the cells from passage 3 (P3) and passage 9 (P9) were utilized to identify MSC cell surface markers and their differentiation capacity. The P3, P5, P7, and P9 cells were used for RNA extraction to qualify the expression of the main immunomodulatory cytokines IDO, VCAM-1, TGF-β, IL-6, IL-10, and PGE2 at mRNA levels. The results indicate that VCAM-1 expression in the subcultures was reduced in bone marrow-derived MSCs. After an increase in P5, P7, and P9, IL-6 expression was reduced. In adipose-derived MSCs, the mRNA levels of IL-10 in higher passages were decreased compared with P3; other studied cytokines had no significant changes in their expression levels in either bone marrow or adipose-derived MSCs. Based on these results, it can be concluded that a suitable source for MSCs in cell therapy with stable expression of main cytokines, even in higher subcultures, appears to be adipose-derived MSCs with the exception of IL-10 secretion.

摘要

间充质干细胞 (MSCs) 在多种疾病中具有治疗潜力;然而,其使用的安全性和有效性仍存在不确定性。由于其免疫调节特性和缺乏免疫反应性,MSCs 的临床应用正在受到深入研究。因此,对于不同微环境中细胞因子分泌的调节的确切机制有一个清晰的认识似乎至关重要。

在本研究中,采集了人骨髓和脂肪组织样本,并分离和培养外周血单核细胞 (PBMCs)。在 MSC 扩增后,使用第 3 代 (P3) 和第 9 代 (P9) 的细胞来鉴定 MSC 细胞表面标志物及其分化能力。使用 P3、P5、P7 和 P9 细胞进行 RNA 提取,以确定主要免疫调节细胞因子 IDO、VCAM-1、TGF-β、IL-6、IL-10 和 PGE2 在 mRNA 水平上的表达。

结果表明,骨髓来源的 MSC 中 VCAM-1 的表达在亚培养中减少。在 P5、P7 和 P9 增加后,IL-6 的表达减少。在脂肪来源的 MSC 中,与 P3 相比,较高代次的 IL-10 的 mRNA 水平降低;在骨髓或脂肪来源的 MSC 中,其他研究的细胞因子的表达水平没有明显变化。

基于这些结果,可以得出结论,对于细胞治疗中具有稳定表达主要细胞因子的合适 MSC 来源,除了 IL-10 分泌外,似乎是脂肪来源的 MSC。

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