Perfusion increases cell content and matrix synthesis in chondrocyte three-dimensional cultures
Perfusion increases cell content and matrix synthesis in chondrocyte three-dimensional cultures
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DOI：
10.1089/10763270260424169
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发表时间：
2002-10-01
影响因子：
--
通讯作者：
Ratcliffe, A
中科院分区：
文献类型：
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作者：
Davisson, T;Sah, RL;Ratcliffe, A
这项工作研究了灌注对聚乙醇酸 (PGA) 支架上培养的绵羊关节软骨细胞的细胞含量和硫酸化糖胺聚糖合成的影响。将绵羊软骨细胞接种到支架上并培养长达 9 天。在此期间，细胞以高达 170 μum/s 的速度进行灌注。样品用 (SO4)-S-35 进行放射性标记，以量化硫酸化糖胺聚糖 (S-GAG) 的整体合成以及构建体中 S-GAG 的保留。还对构建体进行了 DNA 分析，作为细胞含量的测量。培养过程中进行灌注的构建体的 DNA 含量显着高于静态培养的构建体。基质代谢也通过灌注进行调节，这种调节取决于培养持续时间。与静态对照相比，九天的连续灌注使 S-GAG 合成和沉积增加了约 40%。然而，与对照相比，早期时间点（在最初的 3 天培养期间）的灌注抑制了 S-GAG 的合成和保留。这项工作证明了灌注对体外软骨生长的影响，说明了使用灌注来调节组织工程软骨结构的生长，并有可能增强体外组织生长。
This work examines the effect of perfusion on the cell content and sulfated glycosaminoglycan synthesis of ovine articular chondrocytes cultured on polyglycolic acid (PGA) scaffolds. Ovine chondrocytes were seeded onto the scaffolds and cultured for up to 9 days. During this time the cells were subjected to perfusion at velocities of up to 170 muum/s. The samples were radiolabeled with (SO4)-S-35 to quantify the overall synthesis of sulfated glycosaminoglycans (S-GAGs) and the retention of S-GAGs in the construct. The constructs were also analyzed for DNA as a measure of cellular content. Constructs subjected to perfusion during culture had significantly higher DNA contents than those cultured statically. Matrix metabolism was also modulated by perfusion, with this modulation depending on culture duration. Nine days of continuous perfusion increased S-GAG synthesis and deposition by approximately 40% when compared with static controls. However, perfusion at early time points (during the initial 3-day culture period) suppressed the synthesis and retention of S-GAGs when compared with controls. This work demonstrates the effects of perfusion on cartilage growth in vitro, illustrating the use of perfusion to modulate the growth of tissue-engineered cartilage constructs, and potentially enhance tissue growth in vitro.