Transfusion of donor-derived red blood cells (RBCs) is the most common form of cell therapy. Production of transfusion-ready cultured RBCs (cRBCs) is a promising replacement for the current, fully donor-dependent therapy. A single transfusion unit, however, contains 2 × 10¹² RBC, which requires large scale production. Here, we report on the scale-up of cRBC production from static cultures of erythroblasts to 3 L stirred tank bioreactors, and identify the effect of operating conditions on the efficiency of the process. Oxygen requirement of proliferating erythroblasts (0.55–2.01 pg/cell/h) required sparging of air to maintain the dissolved oxygen concentration at the tested setpoint (2.88 mg O₂/L). Erythroblasts could be cultured at dissolved oxygen concentrations as low as 0.7 O₂ mg/ml without negative impact on proliferation, viability or differentiation dynamics. Stirring speeds of up to 600 rpm supported erythroblast proliferation, while 1800 rpm led to a transient halt in growth and accelerated differentiation followed by a recovery after 5 days of culture. Erythroblasts differentiated in bioreactors, with final enucleation levels and hemoglobin content similar to parallel cultures under static conditions.

中文翻译：

---

体外培养的成红细胞在可扩展搅拌生物反应器中的扩增和分化

输注供体来源的红细胞 (RBC) 是最常见的细胞疗法形式。准备输血的培养红细胞 (cRBC) 的生产是目前完全依赖供体的治疗的有前途的替代品。然而，单个输血单位包含 2 × 10 ^{12 个}红细胞，这需要大规模生产。在这里，我们报告了从成红细胞静态培养到 3 L 搅拌罐生物反应器的 cRBC 生产的放大，并确定了操作条件对过程效率的影响。增殖成红细胞的氧气需求 (0.55–2.01 pg/cell/h) 需要喷射空气以将溶解氧浓度维持在测试设定点 (2.88 mg O ₂ /L)。成红细胞可以在低至 0.7 O 的溶解氧浓度下培养₂ mg/ml 对增殖、活力或分化动力学没有负面影响。高达 600 rpm 的搅拌速度支持成红细胞增殖，而 1800 rpm 导致生长暂时停止并加速分化，随后在培养 5 天后恢复。成红细胞在生物反应器中分化，最终去核水平和血红蛋白含量与静态条件下的平行培养相似。