Bioprocess considerations for T-cell therapy: Investigating the impact of agitation, dissolved oxygen, and pH on T-cell expansion and differentiation.
Bioprocess considerations for T-cell therapy: Investigating the impact of agitation, dissolved oxygen, and pH on T-cell expansion and differentiation.
复制标题
T 细胞治疗的生物过程注意事项：研究搅拌、溶解氧和 pH 对 T 细胞扩增和分化的影响。
DOI：
10.1002/bit.27468
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发表时间：
2020
影响因子：
3.8
通讯作者：
Amini A
中科院分区：
文献类型：
--
作者：
Amini A
连续性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，制造工艺的耐用性和再现性仍然是一个挑战。在此，评估了一次性24孔微型生物反应器（微型矩阵）作为高通量筛选工具的用途，以研究不同振荡速度、溶解氧（DO）和pH水平对灌注模拟过程中原代T细胞生长和分化的影响和相互作用。全析因设计允许生成预测模型，其用于找到最佳培养条件。搅动被证明在T细胞的增殖中起着重要作用。200 rpm的振荡速度大大提高了最终活细胞浓度（VCC），而在整个培养过程中，活力保持在90%以上。VCCs最高达9.22 × 106 cells/ml。发现CD 8+中央记忆T细胞（TCM）的分布在很大程度上不受振荡速度的影响。对于细胞生长建立了pH和DO之间的明确相互作用（p< .001），并且确定了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。
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 × 106cells/ml. The distribution of CD8+ central memory T cells (TCM), 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.