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首页> 外文期刊>European Journal of Histochemistry >Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes
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Three-dimensional scaffold-free fusion culture: the way to enhance chondrogenesis of in vitro propagated human articular chondrocytes

机译：三维无支架融合培养：增强体外增殖的人类关节软骨细胞软骨形成的方法

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Cartilage regeneration based on isolated and culture-expanded chondrocytes has been studied in various in vitro models, but the quality varies with respect to the morphology and the physiology of the synthesized tissues. The aim of our study was to promote in vitro chondrogenesis of human articular chondrocytes using a novel three-dimensional (3-D) cultivation system in combination with the chondrogenic differentiation factors transforming growth factor beta 2 (TGF-b2) and L-ascorbic acid. Articular chondrocytes isolated from six elderly patients were expanded in monolayer culture. A single-cell suspension of the dedifferentiated chondrocytes was then added to agar-coated dishes without using any scaffold material, in the presence, or absence of TGF-b2 and/or L-ascorbic acid. Three-dimensional cartilage-like constructs, called single spheroids, and microtissues consisting of several spheroids fused together, named as fusions, were formed. Generated tissues were mainly characterized using histological and immunohistochemical techniques. The morphology of the in vitro tissues shared some similarities to native hyaline cartilage in regard to differentiated S100-positive chondrocytes within a cartilaginous matrix, with strong collagen type II expression and increased synthesis of proteoglycans. Finally, our innovative scaffold-free fusion culture technique supported enhanced chondrogenesis of human articular chondrocytes in vitro . These 3-D hyaline cartilage-like microtissues will be useful for in vitro studies of cartilage differentiation and regeneration, enabling optimization of functional tissue engineering and possibly contributing to the development of new approaches to treat traumatic cartilage defects or osteoarthritis.
机译：已经在各种体外模型中研究了基于分离的和培养扩增的软骨细胞的软骨再生，但是其质量随合成组织的形态和生理学而变化。我们研究的目的是通过使用新型三维（3-D）培养系统以及转化生长因子β2（TGF-b2）和L-抗坏血酸的软骨分化因子来促进人关节软骨细胞的软骨形成。从六名老年患者分离出的关节软骨细胞在单层培养中扩增。然后在存在或不存在TGF-b2和/或L-抗坏血酸的情况下，将未分化的软骨细胞的单细胞悬浮液添加到琼脂包被的皿中，而无需使用任何支架材料。形成了称为单个球体的三维软骨样结构，以及由几个融合在一起的球体组成的微组织，称为融合体。产生的组织主要使用组织学和免疫组织化学技术进行表征。体外组织的形态与天然透明软骨在软骨基质内分化的S100阳性软骨细胞方面具有某些相似之处，其中II型胶原蛋白的表达很强，并且蛋白聚糖的合成增加。最后，我们创新的无支架融合培养技术支持增强人关节软骨细胞的软骨生成。这些3-D透明质酸软骨样微组织将可用于软骨分化和再生的体外研究，从而优化功能组织工程，并可能有助于开发治疗创伤性软骨缺损或骨关节炎的新方法。

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