Ng Ee Xien, Wang Ming, Neo Shu Hui, Tee Ching Ann, Chen Chia-Hung, Van Vliet Krystyn J
Biosystems and Micromechanics, CREATE, Singapore-MIT Alliance for Research and Technology, Singapore, 138602, Singapore.
Critical Analytics for Manufacturing Personalized-Medicine, CREATE, Singapore-MIT Alliance for Research and Technology, Singapore, 138602, Singapore.
Biotechnol J. 2021 Mar;16(3):e2000048. doi: 10.1002/biot.202000048. Epub 2020 Dec 11.
Microcarriers are synthetic particles used in bioreactor-based cell manufacturing of anchorage-dependent cells to promote proliferation at efficient physical volumes, mainly by increasing the surface area-to-volume ratio. Mesenchymal stromal cells (MSCs) are adherent cells that are used for numerous clinical trials of autologous and allogeneic cell therapy, thus requiring avenues for large-scale cell production at efficiently low volumes and cost. Here, a dissolvable gelatin-based microcarrier is developed for MSC expansion. This novel microcarrier shows comparable cell attachment efficiency and proliferation rate when compared to several commercial microcarriers, but with higher harvesting yield due to the direct dissolution of microcarrier particles and thus reduced cell loss at the cell harvesting step. Furthermore, gene expression and in vitro differentiation suggest that MSCs cultured on gelatin microcarriers maintain trilineage differentiation with similar adipogenic differentiation efficiency and higher chondrogenic and osteogenic differentiation efficiency when compared to MSCs cultured on 2D planar polystyrene tissue culture flask; on the contrary, MSCs cultured on conventional microcarriers appear to be bipotent along osteochondral lineages whereby adipogenic differentiation potential is impeded. These results suggest that these gelatin microcarriers are suitable for MSC culture and expansion, and can also potentially be extended for other types of anchorage-dependent cells.
微载体是用于基于生物反应器的贴壁依赖性细胞制造的合成颗粒，主要通过增加表面积与体积比，在有效的物理体积内促进细胞增殖。间充质基质细胞（MSC）是贴壁细胞，用于众多自体和同种异体细胞治疗的临床试验，因此需要以高效的低体积和低成本进行大规模细胞生产的途径。在此，开发了一种用于MSC扩增的可溶解的明胶基微载体。与几种商业微载体相比，这种新型微载体显示出相当的细胞附着效率和增殖率，但由于微载体颗粒的直接溶解，收获产量更高，因此在细胞收获步骤中细胞损失减少。此外，基因表达和体外分化表明，与在二维平面聚苯乙烯组织培养瓶上培养的MSC相比，在明胶微载体上培养的MSC保持三系分化，具有相似的成脂分化效率以及更高的成软骨和成骨分化效率；相反，在传统微载体上培养的MSC似乎沿骨软骨谱系具有双能性，而成脂分化潜能受到阻碍。这些结果表明，这些明胶微载体适用于MSC培养和扩增，并且还可能扩展到其他类型的贴壁依赖性细胞。
