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类风湿关节炎和骨关节炎患者间充质干细胞的软骨形成潜能:微培养系统中的测量

Chondrogenic potential of mesenchymal stem cells from patients with rheumatoid arthritis and osteoarthritis: measurements in a microculture system.

作者信息

Dudics Valeria, Kunstár Aliz, Kovács János, Lakatos Tamás, Géher Pál, Gömör Béla, Monostori Eva, Uher Ferenc

机构信息

Polyclinic of the Hospitaller Brothers of St. John of God, Budapest, Hungary.

出版信息

Cells Tissues Organs. 2009;189(5):307-16. doi: 10.1159/000140679. Epub 2008 Jun 19.

Abstract

BACKGROUND

Mesenchymal stem cells (MSCs) have the potential to differentiate into distinct mesenchymal tissues; including cartilage and bone, they can be an attractive cell source for cartilage tissue engineering approaches. Our objective here was to compare the in vitro chondrogenic potential of MSCs isolated from patients with rheumatoid arthritis (RA) and osteoarthritis (OA) with cells from normal donors.

METHODS

Marrow samples were removed during bone surgery and adherent cell cultures were established. The cells were then passed into a newly developed microaggregate culture system in a medium containing transforming growth factor beta3, insulin, dexamethasone and/or demineralized bone matrix. In vitro chondrogenic activity was measured as metabolic sulfate incorporation and type II collagen expression in pellet cultures.

RESULTS

Culture-expanded MSCs from RA and OA patients did not differ significantly from the normal population with respect to their chondrogenic potential in vitro. Capability of total protein and proteoglycan synthesis as well as collagen II mRNA expression by cell aggregates was similar for all cell preparations in the presence of the appropriate growth and differentiation factors. Chondroprotective drugs such as chondroitin sulfate and glucosamine enhanced, whereas chloroquine inhibited chondrogenesis in normal donor-derived or patient-derived MSC cultures. Galectin-1, a beta-galactoside-binding protein with marked anti-inflammatory activity, stimulated the chondrogenic differentiation of mesenchymal cells in low (<2 microg/ml) concentration.

DISCUSSION

These findings show that MSCs from RA and OA patients possess similar chondrogenic potential as MSCs isolated from healthy donors, therefore these cells may serve as a potential new prospect in cartilage replacement therapy.

摘要

背景

间充质干细胞(MSCs)有分化为不同间充质组织的潜力,包括软骨和骨组织,它们可能是软骨组织工程方法中一种有吸引力的细胞来源。我们在此的目的是比较从类风湿性关节炎(RA)和骨关节炎(OA)患者中分离出的MSCs与正常供体来源细胞的体外软骨形成潜力。

方法

在骨手术过程中获取骨髓样本并建立贴壁细胞培养。然后将细胞接种到一种新开发的微聚集体培养系统中,该系统使用含有转化生长因子β3、胰岛素、地塞米松和/或脱矿骨基质的培养基。通过测量沉淀培养物中的代谢性硫酸盐掺入和II型胶原蛋白表达来检测体外软骨形成活性。

结果

来自RA和OA患者的培养扩增MSCs在体外软骨形成潜力方面与正常人群相比无显著差异。在存在适当的生长和分化因子的情况下,所有细胞制剂的细胞聚集体合成总蛋白和蛋白聚糖的能力以及胶原蛋白II mRNA表达相似。硫酸软骨素和氨基葡萄糖等软骨保护药物可增强正常供体来源或患者来源的MSC培养物中的软骨形成,而氯喹则抑制其软骨形成。半乳糖凝集素-1是一种具有显著抗炎活性的β-半乳糖苷结合蛋白,在低浓度(<2微克/毫升)时可刺激间充质细胞的软骨形成分化。

讨论

这些发现表明,RA和OA患者的MSCs与从健康供体分离出的MSCs具有相似的软骨形成潜力,因此这些细胞可能成为软骨替代治疗的潜在新前景。

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