Vinod Elizabeth, Parameswaran Roshni, Manickam Amirtham Soosai, Livingston Abel, Ramasamy Boopalan, Kachroo Upasana
Department of Physiology, Christian Medical College, Vellore, India.
Centre for Stem Cell Research, Christian Medical College, Vellore, India.
Connect Tissue Res. 2021 Jul;62(4):427-435. doi: 10.1080/03008207.2020.1761344. Epub 2020 May 14.
: Cartilage repair following trauma or degeneration is poor, making cell-based therapy an important avenue of treatment. Chondrocytes and mesenchymal stem cells have been extensively studied as potential candidates, although tendency toward hypertrophy and formation of mixed hyaline-fibrocartilage necessitates further optimization. Chondroprogenitors, isolated using fibronectin adhesion assay are reported to show reduced hypertrophy and enhanced chondrogenesis. Laminin, an essential component of extracellular matrix, has been shown to positively modulate chondrocyte proliferation, migration, and survival. The aim of our study was to evaluate the effect of laminin as a differential adhesion assay and obtain an enriched population of chondroprogenitors and assess its efficiency when compared to progenitors obtained via fibronectin.: Chondrocytes were isolated from three osteoarthritic knee joints and subjected to fibronectin and laminin adhesion to obtain chondroprogenitors. After expansion in culture, they were assessed for differences in their biological characteristics based on growth kinetics, surface marker expression, gene expression for assessing markers of chondrogenesis and hypertrophy, and potential for tri-lineage differentiation.: Our results showed that cells isolated by laminin and fibronectin both displayed comparable characteristics except in terms of proliferative potential (higher in laminin), gene expression of COL2A1 (lower in laminin) and trilineage potential where the laminin group showed higher osteogenic and adipogenic differentiation.: This was the first attempt to successfully isolate human articular cartilage derived chondroprogenitor clones using laminin, which retained stem cell like characteristics. Further evaluation to optimize this method will help enhance chondroprogenitor characteristics, for use in cartilage repair.
创伤或退变后的软骨修复效果不佳,这使得基于细胞的治疗成为重要的治疗途径。软骨细胞和间充质干细胞作为潜在的候选细胞已被广泛研究,尽管存在肥大倾向以及混合透明软骨-纤维软骨的形成,仍需要进一步优化。据报道,使用纤连蛋白黏附试验分离的软骨祖细胞显示出肥大减少和成软骨能力增强。层粘连蛋白是细胞外基质的重要组成部分,已被证明能正向调节软骨细胞的增殖、迁移和存活。我们研究的目的是评估层粘连蛋白作为一种差异黏附试验的效果,获得富集的软骨祖细胞群体,并与通过纤连蛋白获得的祖细胞相比评估其效率。
从三个骨关节炎膝关节分离软骨细胞,使其与纤连蛋白和层粘连蛋白黏附以获得软骨祖细胞。在培养中扩增后,根据生长动力学、表面标志物表达、用于评估成软骨和肥大标志物的基因表达以及三系分化潜能,评估它们生物学特性的差异。
我们的结果表明,通过层粘连蛋白和纤连蛋白分离的细胞除了在增殖潜能(层粘连蛋白组更高)、COL2A1基因表达(层粘连蛋白组更低)以及三系潜能方面(层粘连蛋白组显示出更高的成骨和成脂分化)外,都表现出相似的特征。
这是首次尝试使用层粘连蛋白成功分离出保留干细胞样特征的人关节软骨来源的软骨祖细胞克隆。进一步评估以优化该方法将有助于增强软骨祖细胞特性,用于软骨修复。