Sart Sébastien, Agathos Spiros N
Laboratory of Hydrodynamics (LadHyX) - Department of Mechanics, Ecole Polytechnique, CNRS-UMR7646, 91128, Palaiseau, France.
School of Life Sciences and Biotechnology, Yachay Tech University, 100119, San Miguel de Urcuquí, Ecuador.
Methods Mol Biol. 2016;1502:87-102. doi: 10.1007/7651_2015_314.
Mesenchymal stem cells (MSCs) have emerged as an important tool for tissue engineering, thanks to their differentiation potential and their broad trophic activities. However, for clinical purposes or for relevant in vitro applications, large quantities of MSCs are required, which could hardly be reached using conventional cultivation in plastic dishes. Microcarriers have high surface to volume ratio, which enables the easy scale-up of the expansion and differentiation of MSCs. In addition, the agitation in stirred tank bioreactors limits the diffusion gradient of nutrients or morphogens, thus providing a physiologically relevant environment to favor MSC production at large scale. This work describes a simple method for the mass expansion and differentiation of MSCs, including the procedures to monitor the proliferation, metabolic status and phenotype of MSCs during suspension culture. Moreover, this work proposes suitable materials for cGMP compliant culture conditions enabling the clinical grade production of MSCs.
间充质干细胞（MSCs）因其分化潜能和广泛的营养活性，已成为组织工程的重要工具。然而，对于临床应用或相关的体外应用，需要大量的间充质干细胞，而使用塑料培养皿中的传统培养方法很难获得所需数量。微载体具有高的表面积与体积比，这使得间充质干细胞的扩增和分化易于放大。此外，搅拌罐生物反应器中的搅拌限制了营养物质或形态发生素的扩散梯度，从而提供了有利于大规模生产间充质干细胞的生理相关环境。这项工作描述了一种间充质干细胞大规模扩增和分化的简单方法，包括在悬浮培养过程中监测间充质干细胞增殖、代谢状态和表型的程序。此外，这项工作提出了适合符合cGMP标准培养条件的材料，能够实现间充质干细胞的临床级生产。
