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Protection of human induced pluripotent stem cells against shear stress in suspension culture by Bingham plastic fluid

机译:用宾厄姆塑料流体保护人诱导多能干细胞对悬浮培养的剪切应力

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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.
机译:悬浮培养是多能干细胞(PSCs)工业制剂的重要方法,用于再生治疗和药物筛选。通常,悬浮培养需要搅拌以保持PSC汇集悬浮并促进传质,但搅拌也会导致细胞损伤。在本研究中,我们研究了使用补充有基于多糖的聚合物的Bingham塑料流体,以保护PSC免受悬浮培养物的细胞损伤。流变分析表明,在用聚合物FP003补充后,培养基获得屈服应力并成为宾果塑料流体。生长/死亡分析显示,在宾厄姆塑料介质中2天的聚集体形成和2天的悬浮液改善了细胞生长和预防细胞死亡。在初始聚集步骤之后,虽然常规培养基的强烈搅拌(120rpm)导致含重的细胞死亡,但在弯曲的塑料流体中,我们获得了与最佳搅拌(90rpm)的正常培养相同的生长。这表明宾厄姆塑料流体保护来自悬浮培养中的剪切应力的细胞,并且可用于在发展大规模时增强其鲁棒性。

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