Laboratory for Reconstructive Plastic Surgery, Department of Clinical and Experimental Medicine, Linköping University, Sweden.
Biomed Mater. 2011 Dec;6(6):065001. doi: 10.1088/1748-6041/6/6/065001. Epub 2011 Sep 29.
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.
本研究考察了四种不同制造工艺的大孔明胶微载体（CultiSpher）对人软骨细胞扩增的影响——初始制备过程中使用的乳化剂的量和明胶交联介质。来自三个供体的单层扩增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。每隔一天提取样本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进行分析。所有微载体的软骨细胞密度均呈线性增加，表明存在持续但有限的细胞增殖。在初始扩增阶段观察到强烈的供体软骨细胞效应。微载体之间的最终细胞产量存在显著差异，我们的结果表明制造差异在这一点上影响了软骨细胞密度。剩余的细胞对软骨形成标记物 SOX-9 和 S-100 呈阳性染色，但细胞外基质形成适度至无法检测。总之，四种明胶微载体在两周的时间内支持软骨细胞的黏附和增殖。观察到低乳化剂含量和在水和丙酮中交联的微载体的最佳产量。这些结果有助于确定特定细胞过程和群体的最佳生物材料参数。
