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首页> 外文期刊>Cell and Tissue Research >Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes
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Effects of oxygen and culture system on in vitro propagation and redifferentiation of osteoarthritic human articular chondrocytes

机译：氧气和培养系统对骨关节炎人关节软骨细胞体外繁殖和再分化的影响

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Regenerative medicine-based approaches for the repair of damaged cartilage rely on the ability to propagate cells while promoting their chondrogenic potential. Thus, conditions for cell expansion should be optimized through careful environmental control. Appropriate oxygen tension and cell expansion substrates and controllable bioreactor systems are probably critical for expansion and subsequent tissue formation during chondrogenic differentiation. Wetherefore evaluated the effects of oxygen and microcarrier culture on the expansion and subsequent differentiation of human osteoarthritic chondrocytes. Freshly isolated chondrocytes were expanded on tissue culture plastic or CultiSpher-G microcarriers under hypoxic or normoxic conditions (5% or 20% oxygen partial pressure, respectively) followed by cell phenotype analysis with flow cytometry. Cells were redifferentiated in micromass pellet cultures over 4 weeks, under either hypoxia or normoxia. Chondrocytes cultured on tissue culture plastic proliferated faster, expressed higher levels of cell surface markers CD44 and CD105 and demonstrated stronger staining for proteoglycans and collagen type II in pellet cultures compared with microcarrier-cultivated cells. Pellet wet weight, glycosaminoglycan content and expression of chondrogenic genes were significantly increased in cells differentiated under hypoxia. Hypoxia-inducible factor- 3α mRNA was up-regulated in these cultures in response to low oxygen tension. These data confirm the beneficial influence of reduced oxygen on ex vivo chondrogenesis. However, hypoxia during cell expansion and microcarrier bioreactor culture does not enhance intrinsic chondrogenic potential. Further improvements in cell culture conditions are therefore required before chondrocytes from osteoarthritic and aged patients can become a useful cell source for cartilage regeneration.
机译：修复受损软骨的基于再生医学的方法依赖于在促进细胞成软骨潜力的同时繁殖细胞的能力。因此，应通过仔细的环境控制来优化细胞扩增的条件。合适的氧气张力和细胞扩增底物以及可控的生物反应器系统可能对软骨分化过程中的扩张和随后的组织形成至关重要。因此，我们评估了氧气和微载体培养对人骨关节炎软骨细胞扩增和随后分化的影响。将新鲜分离的软骨细胞在缺氧或常氧条件下（分别为5％或20％氧分压）在组织培养塑料或CultiSpher-G微载体上扩增，然后通过流式细胞仪进行细胞表型分析。在缺氧或常氧下，经过4周的微团沉淀培养细胞再分化。与微载体培养的细胞相比，在组织培养塑料上培养的软骨细胞增殖更快，表达的细胞表面标志物CD44和CD105含量更高，并且蛋白沉淀和II型胶原蛋白染色更强。在缺氧条件下分化的细胞中，团粒湿重，糖胺聚糖含量和软骨生成基因的表达显着增加。在这些培养物中，低氧诱导因子-3αmRNA的表达上调是由于低氧张力引起的。这些数据证实了减少的氧气对离体软骨形成的有益影响。但是，细胞扩增和微载体生物反应器培养过程中的缺氧不能增强内在的软骨形成潜能。因此，在骨关节炎和老年患者的软骨细胞成为软骨再生的有用细胞来源之前，需要进一步改善细胞培养条件。

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