High-throughput proliferation and activation of NK-92MI cell spheroids via a homemade one-step closed bioreactor in pseudostatic cultures for immunocellular therapy.

IF 5.7 3区 生物学 Q1 BIOCHEMICAL RESEARCH METHODS
Jhih-Ni Lin, Che-Yung Kuan, Chia-Ting Chang, Zhi-Yu Chen, Wei-Ting Kuo, Jason Lin, Yu-Ying Lin, I-Hsuan Yang, Feng-Huei Lin
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Abstract

The NK-92MI cell line has displayed significant promise in clinical trials for cancer treatment. However, challenges persist in obtaining sufficient cell quantities and achieving optimal cytotoxicity. The proliferation of natural killer (NK) cells involves the formation of cell aggregates, but excessively large aggregates can impede nutrient and waste transport, leading to reduced cell survival rates. In this study, a custom bioreactor was designed to mimic pseudostatic culture conditions by integrating brief mechanical rotation during a 6-h static culture period. This method aimed to achieve an optimal aggregate size while improving cell viability. The findings revealed a 144-fold expansion of 3D NK-92MI cell aggregates, reaching an ideal size of 80-150 µm, significantly increasing both cell proliferation and survival rates. After 14 days of culture, the NK-92MI cells maintained their phenotype during the subsequent phase of cell activation. Moreover, these cells presented elevated levels of IFN-γ expression after IL-18 activation, resulting in enhanced NK cell-mediated cytotoxicity against K562 cells. This innovative strategy, which uses a closed suspension-based culture system, presents a promising approach for improving cell expansion and activation techniques in immunocellular therapy.

通过自制的假静态培养一步式封闭生物反应器高通量增殖和激活 NK-92MI 细胞球,用于免疫细胞疗法。
在癌症治疗的临床试验中,NK-92MI 细胞系显示出了巨大的前景。然而,在获得足够数量的细胞和实现最佳细胞毒性方面仍然存在挑战。自然杀伤(NK)细胞的增殖涉及细胞聚集体的形成,但过大的聚集体会阻碍营养物质和废物的运输,导致细胞存活率降低。本研究设计了一种定制生物反应器,通过在 6 小时静态培养期间结合短暂的机械旋转来模拟假静态培养条件。这种方法的目的是在提高细胞存活率的同时达到最佳的聚合大小。研究结果表明,三维 NK-92MI 细胞聚集体扩大了 144 倍,达到了 80-150 微米的理想尺寸,显著提高了细胞增殖率和存活率。培养 14 天后,NK-92MI 细胞在随后的细胞活化阶段保持了其表型。此外,IL-18 激活后,这些细胞的 IFN-γ 表达水平升高,从而增强了 NK 细胞介导的对 K562 细胞的细胞毒性。这种采用封闭悬浮培养系统的创新策略为改进免疫细胞疗法中的细胞扩增和活化技术提供了一种前景广阔的方法。
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来源期刊
Journal of Biological Engineering
Journal of Biological Engineering BIOCHEMICAL RESEARCH METHODS-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
CiteScore
7.10
自引率
1.80%
发文量
32
审稿时长
17 weeks
期刊介绍: Biological engineering is an emerging discipline that encompasses engineering theory and practice connected to and derived from the science of biology, just as mechanical engineering and electrical engineering are rooted in physics and chemical engineering in chemistry. Topical areas include, but are not limited to: Synthetic biology and cellular design Biomolecular, cellular and tissue engineering Bioproduction and metabolic engineering Biosensors Ecological and environmental engineering Biological engineering education and the biodesign process As the official journal of the Institute of Biological Engineering, Journal of Biological Engineering provides a home for the continuum from biological information science, molecules and cells, product formation, wastes and remediation, and educational advances in curriculum content and pedagogy at the undergraduate and graduate-levels. Manuscripts should explore commonalities with other fields of application by providing some discussion of the broader context of the work and how it connects to other areas within the field.
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