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滑膜间充质基质细胞的分离和体外扩增用于软骨修复。

Isolation and ex vivo expansion of synovial mesenchymal stromal cells for cartilage repair.

机构信息

Department of Bioengineering and Institute for Biotechnology and Bioengineering, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.

Centro Hospitalar de Lisboa Ocidental, E.P.E, Hospital São Francisco Xavier, Lisboa, Portugal.

出版信息

Cytotherapy. 2014 Apr;16(4):440-53. doi: 10.1016/j.jcyt.2013.10.010. Epub 2013 Dec 22.

Abstract

BACKGROUND AIMS

Hyaline articular cartilage is a highly specialized tissue that offers a low-friction and wear-resistant interface for weight-bearing surface articulation in diarthrodial joints, but it lacks vascularity. It displays an inherent inability to heal when injured in a skeletally mature individual. Joint-preserving treatment procedures such as mosaicplasty, débridement, perichondrium transplantation and autologous chondrocyte implantation have shown variable results, and the average long-term result is sub-standard. Because of these limitations of the treatment methods and lack of intrinsic repair capacity of mature cartilage tissue, an alternative treatment approach is needed, and synovial mesenchymal stromal cells (SMSCs) represent an attractive therapeutic alternative because of their ex vivo proliferation capacity, multipotency and ability to undergo chondrogenesis.

METHODS

SMSCs were isolated from tissues obtained by arthroscopy using two types of biopsies. Ex vivo cell expansion was accomplished under static and dynamic culture followed by characterization of cells according to the International Society for Cellular Therapy guidelines. Kinetic growth models and metabolite analysis were used for understanding the growth profile of these cells.

RESULTS

For the first time, SMSCs were expanded in stirred bioreactors and achieved higher cell density in a shorter period of time compared with static culture or with other mesenchymal stromal cell sources.

CONCLUSIONS

In this study we were able to achieve (8.8 ± 0.2) × 10(5) cells within <2 weeks in dynamic culture under complete xeno-free conditions. Our results also provided evidence that after dynamic culture these cells had an up-regulation of chondrogenic genes, which can be a potential factor for articular cartilage regeneration in clinical settings.

摘要

背景目的

透明关节软骨是一种高度特化的组织,为负重关节的关节面提供低摩擦和耐磨的界面,但它缺乏血管。在骨骼成熟的个体中,受伤后它显示出无法自行愈合的固有能力。关节保存治疗程序,如马赛克plasty、清创术、软骨膜移植和自体软骨细胞植入,已经显示出不同的结果,平均长期结果是低于标准的。由于这些治疗方法的局限性和成熟软骨组织缺乏内在修复能力,需要一种替代的治疗方法,滑膜间充质基质细胞(SMSCs)由于其体外增殖能力、多能性和软骨形成能力,成为一种有吸引力的治疗选择。

方法

使用两种类型的活检,从关节镜获得的组织中分离 SMSCs。通过静态和动态培养进行体外细胞扩增,然后根据国际细胞治疗学会的指南对细胞进行表征。使用动力学生长模型和代谢物分析来了解这些细胞的生长情况。

结果

首次在搅拌生物反应器中扩增 SMSCs,并与静态培养或其他间充质基质细胞来源相比,在更短的时间内实现更高的细胞密度。

结论

在这项研究中,我们能够在完全无动物来源的条件下,在动态培养中在不到 2 周的时间内达到(8.8±0.2)×10(5)个细胞。我们的结果还提供了证据,表明这些细胞在动态培养后,软骨生成基因的表达上调,这可能是临床环境中关节软骨再生的一个潜在因素。

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