Graduate Program in Experimental Biology, DCBS, Universidad Autónoma Metropolitana Iztapalapa, Mexico City 09340, Mexico.
Laboratorio de Gerociencias, Instituto Nacional de Rehabilitación "Luis Guillermo Ibarra Ibara", Mexico City 14389, Mexico.
Cells. 2022 Aug 17;11(16):2553. doi: 10.3390/cells11162553.
The preservation of the chondrogenic phenotype and hypoxia-related physiological microenvironment are major challenges in the 2D culture of primary human chondrocytes. To address this problem, we develop a 3D culture system generating scaffold-free spheroids from human chondrocytes. Our results highlight the chondrogenic potential of cultured human articular chondrocytes in a 3D system combined with hypoxia independently of the cartilage source. After 14 days of culture, we developed spheroids with homogenous diameter and shape from hyaline cartilage donors. Spheroids generated in hypoxia showed a significantly increased glycosaminoglycans synthesis and up-regulated the expression of , , , , and compared to those obtained under normoxic conditions. Therefore, we conclude that spheroids developed under hypoxic conditions modulate the expression of chondrogenesis-related genes and native tissue features better than 2D cultures. Thus, this scaffold-free 3D culture system represents a novel in vitro model that can be used for cartilage biology research.
在二维培养原代人软骨细胞的过程中，维持软骨细胞表型和缺氧相关的生理微环境是主要挑战。为了解决这个问题，我们开发了一种 3D 培养系统，可从人软骨细胞生成无支架的球体。我们的结果突出显示了在 3D 系统中培养的人关节软骨细胞在缺氧条件下的软骨生成潜力，而与软骨来源无关。培养 14 天后，我们从透明软骨供体中开发出具有均匀直径和形状的球体。与在常氧条件下获得的球体相比，在低氧条件下生成的球体的糖胺聚糖合成显著增加，并且上调了 、 、 、 和 的表达。因此，我们得出结论，缺氧条件下形成的球体比 2D 培养更好地调节软骨生成相关基因和天然组织特征的表达。因此，这种无支架 3D 培养系统代表了一种可用于软骨生物学研究的新型体外模型。
