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人脐带间充质干细胞提高 OA 退变软骨细胞的活力。

The human umbilical cord stem cells improve the viability of OA degenerated chondrocytes.

机构信息

Teaching Center of Experimental Medicine, Shanghai Medical College, Fudan University, Shanghai 200032, P.R. China.

Department of Orthopedics, 455th Hospital of PLA, Shanghai 200052, P.R. China.

出版信息

Mol Med Rep. 2018 Mar;17(3):4474-4482. doi: 10.3892/mmr.2018.8413. Epub 2018 Jan 9.

Abstract

Osteoarthritis (OA) affects a large number of patients; however, human umbilical cord stem cells exhibit therapeutic potential for treating OA. The aim of the present study was to explore the interaction between human umbilical cord stem cells and degenerated chondrocytes, and the therapeutic potential of human umbilical cord stem cells on degenerated chondrocytes. Human umbilical cord‑derived mesenchymal stem cells (hUC‑MSCs) were harvested from human umbilical cords, and flow cytometry was used to analyze the surface antigen markers, in addition, chondrogenic, osteogenic and adipogenic differentiation on the cells was investigated. OA cells at P3 were cocultured with hUC‑MSCs in a separated co‑culture system, and reverse transcription‑polymerase chain reaction and western blot were used to evaluate the mRNA, and protein expression of collagen type II (Col2), SRY‑box 9 (sox‑9) and aggrecan. The level of inflammatory cytokines, tumor necrosis factor‑α, interleukin (IL)‑1β, IL‑6, IL‑10, were analyzed by ELISA in the supernatant. hUC‑MSCs grow in a fibroblastic shape with stable proliferation. hUC‑MSCs expressed cluster of differentiation 44 (CD44), CD73, CD90, CD105; while did not express CD34, CD45, CD106, CD133. After multi‑induction, hUC‑MSCs were able to differatiate into adipogenic, osteogenic and chondrogenic lineage. hUC‑MSCs inhibited the expression of matrix metalloproteinase‑13, collagen type X α1 chain and cyclooxygenase‑2 in OA chondrocytes, and enhanced the proliferation of OA chondrocytes, while OA chondrocytes stimulated the production of Col2, sox‑9 and aggrecan and promoted hUC‑MSCs differentiate into chondrocytes. Flow cytometry analysis demonstrated hUC‑MSCs have a predominant expression of stem cell markers, while the hematopoietic and endothelial markers were absent. Osteogenic, chondrogenic and adipogenic differentiation was observed in certain induction conditions. hUC‑MSCs improved the proliferation of OA chondrocytes and downregulated the expression of inflammatory cytokines, while OA chondrocytes promoted MSCs to differentiate into chondrocytes. Taken together, the co‑culture of hUC‑MSCs and OA chondrocytes may provide a therapeutic potential in OA treatment.

摘要

骨关节炎(OA)影响大量患者;然而,人脐带干细胞对治疗 OA 具有治疗潜力。本研究旨在探讨人脐带干细胞与退变软骨细胞的相互作用,以及人脐带干细胞对退变软骨细胞的治疗潜力。从人脐带中提取人脐带间充质干细胞(hUC-MSCs),并通过流式细胞术分析细胞表面抗原标志物,此外,还研究了细胞的成软骨、成骨和成脂分化。在分离的共培养系统中,将第 3 代 OA 细胞与 hUC-MSCs 共培养,通过逆转录-聚合酶链反应和蛋白质印迹法评估胶原 II(Col2)、性别决定区 Y 框 9(sox-9)和聚集蛋白聚糖的 mRNA 和蛋白表达。通过酶联免疫吸附试验(ELISA)分析上清液中炎症细胞因子肿瘤坏死因子-α、白细胞介素(IL)-1β、IL-6 和 IL-10 的水平。hUC-MSCs 呈成纤维细胞样生长,增殖稳定。hUC-MSCs 表达 CD44、CD73、CD90、CD105;而不表达 CD34、CD45、CD106、CD133。经多诱导后,hUC-MSCs 可分化为成脂、成骨和成软骨谱系。hUC-MSCs 抑制 OA 软骨细胞中基质金属蛋白酶-13、胶原 X 链α1 链和环氧化酶-2 的表达,促进 OA 软骨细胞的增殖,而 OA 软骨细胞刺激 Col2、sox-9 和聚集蛋白聚糖的产生,并促进 hUC-MSCs 向软骨细胞分化。流式细胞术分析表明,hUC-MSCs 具有较高的干细胞标志物表达,而造血和内皮标志物则不存在。在某些诱导条件下观察到成骨、成软骨和成脂分化。hUC-MSCs 改善 OA 软骨细胞的增殖,下调炎症细胞因子的表达,而 OA 软骨细胞促进 MSC 向软骨细胞分化。综上所述,hUC-MSCs 与 OA 软骨细胞的共培养可能为 OA 的治疗提供治疗潜力。

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