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Growth factors for clinical-scale expansion of human articular chondrocytes : Relevance for automated bioreactor systems

机译:人类关节软骨细胞临床规模扩张的生长因子:自动生物反应器系统的相关性

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摘要

The expansion of chondrocytes in automated bioreactors for clinical use requires that a relevant number of cells be generated, starting from variable initial seeding densities in one passage and using autologous serum. We investigated whether the growth factor combination transforming growth factor beta 1/fibroblast growth factor 2/platelet-derived growth factor BB (TFP), recently shown to enhance the proliferation capacity of human articular chondrocytes (HACs), allows the efficiency of chondrocyte use to be increased at different seeding densities and percentages of human serum (HS). HACs were seeded at 1,000, 5,000, and 10,000 celIS/cm(2) in medium containing 10 bovine serum or 10,000 cells/cm(2) with 1 chondrogenic capacity of post-expanded HACs was then assessed in pellet cultures. Expansion with TFP allowed a sufficient number of HACs to be obtained in one passage even at the lowest seeding density and HS percentage and variability in cartilage-forming capacity of HACs expanded under the different conditions to be reduced. Instead, larger variations and insufficient yields were found in the absence of TFP. By allowing large numbers of cells to be obtained, starting from a wide range of initial seeding densities and HS percentages, the use of TFP may represent a viable solution for the efficient expansion of HACs and addresses constraints of automated clinical bioreactor systems.
机译:用于临床用途的自动生物反应器中软骨细胞的扩增需要产生一定数量的细胞,从一次传代中可变的初始接种密度开始,并使用自体血清。我们调查了生长因子组合转化生长因子β1/成纤维细胞生长因子2 /血小板衍生的生长因子BB(TFP)是否最近能增强人关节软骨细胞(HACs)的增殖能力,从而使软骨细胞的利用效率达到在不同的播种密度和人血清(HS)百分比下增加。将HAC分别以1,000、5,000和10,000 celIS / cm(2)的浓度接种在含有10个牛血清或10,000个细胞/ cm(2)的培养基中,其中1种软骨具有扩建后的HAC的成软骨能力,然后在沉淀培养物中进行评估。使用TFP进行扩增,即使在最低播种密度和HS百分比下,也可以通过一次传代获得足够数量的HAC,并且可以降低在不同条件下扩增的HAC的软骨形成能力的差异。取而代之的是,在没有TFP的情况下发现了较大的变异和不足的产量。通过允许从大量的初始接种密度和HS百分比范围开始获得大量细胞,TFP的使用可能代表了有效扩展HAC的可行解决方案,并解决了自动化临床生物反应器系统的局限性。

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