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成年小鼠半月板干/祖细胞的鉴定与特性分析

Identification and characterization of adult mouse meniscus stem/progenitor cells.

作者信息

Gamer Laura W, Shi Rui Rui, Gendelman Ashira, Mathewson Dylan, Gamer Jackson, Rosen Vicki

机构信息

a Department of Developmental Biology , Harvard School of Dental Medicine , Boston , MA , USA.

出版信息

Connect Tissue Res. 2017 May-Jul;58(3-4):238-245. doi: 10.1080/03008207.2016.1271797. Epub 2016 Dec 22.

Abstract

Meniscal damage is a common problem that accelerates the onset of knee osteoarthritis. Stem cell-based tissue engineering treatment approaches have shown promise in preserving meniscal tissue and restoring meniscal function. The purpose of our study was to identify meniscus-derived stem/progenitor cells (MSPCs) from mouse, a model system that allows for in vivo analysis of the mechanisms underlying meniscal injury and healing. MSPCs were isolated from murine menisci grown in explant culture and characterized for stem cell properties. Flow cytometry was used to detect the presence of surface antigens related to stem cells, and qRT-PCR was used to examine the gene expression profile of MSPCs. Major proteins associated with MSPCs were localized in the adult mouse knee using immunohistochemistry. Our data show that MSPCs have universal stem cell-like properties including clonogenicity and multi-potentiality. MSPCs expressed the mesenchymal stem cell markers CD44, Sca-1, CD90, and CD73 and when cultured had elevated levels of biglycan and collagen type I, important extracellular matrix components of adult meniscus. MSPC also expressed significant levels of Lox and Igf-1, genes associated with the embryonic meniscus. Localization studies showed staining for these same proteins in the superficial and outer zones of the adult mouse meniscus, regions thought to harbor endogenous repair cells. MSPCs represent a novel resident stem cell population in the murine meniscus. Analysis of MSPCs in mice will allow for a greater understanding of the cell biology of the meniscus, essential information for enhancing therapeutic strategies for treating knee joint injury and disease.

摘要

半月板损伤是加速膝关节骨关节炎发病的常见问题。基于干细胞的组织工程治疗方法在保留半月板组织和恢复半月板功能方面已显示出前景。我们研究的目的是从小鼠中鉴定半月板来源的干/祖细胞(MSPCs),小鼠是一种可用于体内分析半月板损伤和愈合潜在机制的模型系统。MSPCs从外植体培养中生长的小鼠半月板中分离出来,并对其干细胞特性进行表征。流式细胞术用于检测与干细胞相关的表面抗原的存在,qRT-PCR用于检测MSPCs的基因表达谱。使用免疫组织化学将与MSPCs相关的主要蛋白定位在成年小鼠膝关节中。我们的数据表明,MSPCs具有包括克隆形成能力和多能性在内的普遍干细胞样特性。MSPCs表达间充质干细胞标志物CD44、Sca-1、CD90和CD73,培养时其双糖链蛋白聚糖和I型胶原蛋白水平升高,这两种蛋白是成年半月板重要的细胞外基质成分。MSPCs还表达了与胚胎半月板相关的基因Lox和Igf-1的显著水平。定位研究显示,成年小鼠半月板的表层和外层区域存在这些相同蛋白的染色,这些区域被认为含有内源性修复细胞。MSPCs代表了小鼠半月板中一种新的常驻干细胞群体。对小鼠MSPCs的分析将有助于更深入了解半月板的细胞生物学,这是增强膝关节损伤和疾病治疗策略的重要信息。

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