Department of Biotechnology, Osaka University, Osaka, Japan.
Department of Chemical System Engineering, The University of Tokyo, Tokyo, Japan.
Biotechnol Prog. 2021 Mar;37(2):e3100. doi: 10.1002/btpr.3100. Epub 2020 Nov 28.
Suspension culture is an important method used in the industrial preparation of pluripotent stem cells (PSCs), for regenerative therapy and drug screening. Generally, a suspension culture requires agitation to keep PSC aggregates suspended and to promote mass transfer, but agitation also causes cell damage. In this study, we investigated the use of a Bingham plastic fluid, supplemented with a polysaccharide-based polymer, to preserve PSCs from cell damage in suspension culture. Rheometric analysis showed that the culture medium gained yield stress and became a Bingham plastic fluid, after supplementation with the polymer FP003. A growth/death analysis revealed that 2 days of aggregate formation and 2 days of suspension in the Bingham plastic medium improved cell growth and prevented cell death. After the initial aggregation step, whereas strong agitation (120 rpm) of a conventional culture medium resulted in massive cell death, in the Bingham plastic fluid we obtained the same growth as the normal culture with optimal agitation (90 rpm). This indicates that Bingham plastic fluid protected cells from shear stress in suspension culture and could be used to enhance their robustness when developing a large-scale.
悬浮培养是一种在多能干细胞（PSCs）的工业制备中使用的重要方法，用于再生治疗和药物筛选。通常，悬浮培养需要搅拌来保持 PSC 聚集体悬浮并促进传质，但搅拌也会导致细胞损伤。在这项研究中，我们研究了使用宾汉塑性流体，辅以基于多糖的聚合物，来防止 PSCs 在悬浮培养中受到细胞损伤。流变分析表明，在添加聚合物 FP003 后，培养基获得了屈服应力并成为宾汉塑性流体。生长/死亡分析表明，2 天的聚集体形成和 2 天的宾汉塑性培养基悬浮培养可促进细胞生长并防止细胞死亡。在初始聚集步骤之后，虽然传统培养基的强搅拌（120rpm）会导致大量细胞死亡，但在宾汉塑性流体中，我们以最佳搅拌（90rpm）获得了与正常培养相同的生长。这表明宾汉塑性流体在悬浮培养中保护细胞免受剪切应力，并可用于增强大规模培养时的稳健性。
