Adoptive T‐cell therapy (ACT) has emerged as a promising new way to treat systemic cancers such as acute lymphoblastic leukemia. However, the robustness and reproducibility of the manufacturing process remains a challenge. Here, a single‐use 24‐well microbioreactor (micro‐Matrix) was assessed for its use as a high‐throughput screening tool to investigate the effect and the interaction of different shaking speeds, dissolved oxygen (DO), and pH levels on the growth and differentiation of primary T cells in a perfusion‐mimic process. The full factorial design allowed for the generation of predictive models, which were used to find optimal culture conditions. Agitation was shown to play a fundamental role in the proliferation of T cells. A shaking speed of 200 rpm drastically improved the final viable cell concentration (VCC), while the viability was maintained above 90% throughout the cultivation. VCCs reached a maximum of 9.22 × 10⁶ cells/ml. The distribution of CD8+ central memory T cells (T_CM), was found to be largely unaffected by the shaking speed. A clear interaction between pH and DO (p < .001) was established for the cell growth and the optimal culture conditions were identified for a combination of 200 rpm, 25% DO, and pH of 7.4. The combination of microbioreactor technology and Design of Experiment methodology provides a powerful tool to rapidly gain an understanding of the design space of the T‐cell manufacturing process.

中文翻译：

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T 细胞治疗的生物过程注意事项：研究搅拌、溶解氧和 pH 值对 T 细胞扩增和分化的影响。

过继性 T 细胞疗法 (ACT) 已成为治疗急性淋巴细胞白血病等全身性癌症的一种有前途的新方法。然而，制造过程的稳健性和可重复性仍然是一个挑战。在这里，评估了一次性 24 孔微生物反应器 (micro-Matrix) 作为高通量筛选工具的用途，以研究不同摇动速度、溶解氧 (DO) 和 pH 水平对反应的影响和相互作用。灌注模拟过程中原代 T 细胞的生长和分化。全因子设计允许生成预测模型，用于寻找最佳培养条件。研究表明，搅拌在 T 细胞增殖中起着重要作用。200 rpm 的振荡速度极大地提高了最终活细胞浓度 (VCC)，而在整个培养过程中存活率保持在90%以上。VCC 达到最大值 9.22 × 10^{6 个}细胞/毫升。发现 CD8+ 中央记忆 T 细胞 (T _CM ) 的分布在很大程度上不受摇动速度的影响。pH 和 DO 之间存在明显的相互作用 ( p  < .001)，用于细胞生长，并确定了 200 rpm、25% DO 和 pH 7.4 的组合的最佳培养条件。微生物反应器技术和实验设计方法的结合提供了一个强大的工具，可以快速了解 T 细胞制造过程的设计空间。