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Cell expansion of human articular chondrocytes on macroporous gelatine scaffolds - Impact of microcarrier selection on cell proliferation

机译:大孔明胶支架上人关节软骨细胞的细胞扩增-微载体选择对细胞增殖的影响

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This study investigates human chondrocyte expansion on four macroporous gelatine microcarriers (CultiSpher) differing with respect to two manufacturing processes - the amount of emulsifier used during initial preparation and the gelatine cross-linking medium. Monolayer-expanded articular chondrocytes from three donors were seeded onto the microcarriers and cultured in spinner flask systems for a total of 15 days. Samples were extracted every other day to monitor cell viability and establish cell counts, which were analysed using analysis of variance and piecewise linear regression. Chondrocyte densities increased according to a linear pattern for all microcarriers, indicating an ongoing, though limited, cell proliferation. A strong chondrocyte donor effect was seen during the initial expansion phase. The final cell yield differed significantly between the microcarriers and our results indicate that manufacturing differences affected chondrocyte densities at this point. Remaining cells stained positive for chondrogenic markers SOX-9 and S-100 but extracellular matrix formation was modest to undetectable. In conclusion, the four gelatine microcarriers supported chondrocyte adhesion and proliferation over a two week period. The best yield was observed for microcarriers produced with low emulsifier content and cross-linked in water and acetone. These results add to the identification of optimal biomaterial parameters for specific cellular processes and populations.
机译:这项研究调查了人类软骨细胞在四种大孔明胶微载体(CultiSpher)上的扩增,这两种载体在两种制造工艺上有所不同-初始制备过程中使用的乳化剂数量和明胶交联介质。将来自三个供体的单层扩张的关节软骨细胞接种到微载体上,并在旋转瓶系统中培养总共15天。每隔一天提取样品以监测细胞活力并建立细胞计数,使用方差分析和分段线性回归对其进行分析。对于所有微载体,软骨细胞的密度均按照线性模式增加,这表明正在进行的细胞增殖(尽管有限)。在最初的扩张阶段看到了强烈的软骨细胞供体作用。微载体之间的最终细胞产量差异显着,我们的结果表明,此时的制造差异会影响软骨细胞的密度。剩余的细胞对软骨形成标记SOX-9和S-100染色呈阳性,但细胞外基质的形成适中至无法检测。总之,四种明胶微载体在两周内支持软骨细胞粘附和增殖。对于具有低乳化剂含量并在水和丙酮中​​交联的微载体,观察到最佳产率。这些结果增加了针对特定细胞过程和群体的最佳生物材料参数的鉴定。

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