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动态悬浮培养通过上调细胞活化和葡萄糖消耗率来提高细胞因子诱导的杀伤细胞的体外扩增。

Dynamic suspension culture improves ex vivo expansion of cytokine-induced killer cells by upregulating cell activation and glucose consumption rate.

机构信息

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.

State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, PR China.

出版信息

J Biotechnol. 2018 Dec 10;287:8-17. doi: 10.1016/j.jbiotec.2018.09.010. Epub 2018 Sep 29.

Abstract

Ex vivo expansion is an effective strategy to acquire cytokine-induced killer (CIK) cells needed for clinical trials. In this work, the effects of dynamic suspension culture, which was carried out by shake flasks on a shaker, on CIK cells were investigated by the analysis of expansion characteristics and physiological functions, with the objective to optimize the culture conditions for ex vivo expansion of CIK cells. The results showed that the expansion folds of total cells in dynamic cultures reached 69.36 ± 30.36 folds on day 14, which were significantly higher than those in static cultures (9.24 ± 1.12 folds, P < 0.05), however, the proportions of CD3 cells and CD3CD56 cells in both cultures were similar, leading to much higher expansion of CD3 cells and CD3CD56 cells in dynamic cultures. Additionally, expanded CIK cells in two cultures possessed comparable physiological functions. Notably, significantly higher percentages of CD25 cells and CD69 cells were found in dynamic cultures (P <  0.05). Besides, much higher glucose consumption rate of cells (P <  0.05) but similar Y were observed in dynamic cultures. Further, cells in dynamic cultures had better glucose utilization efficiency. Together, these results suggested that dynamic cultures improved cell activation, then accelerated glucose consumption rate, which enhanced cell expansion and promoted glucose utilization efficiency of cells.

摘要

体外扩增是获取临床试验所需细胞因子诱导的杀伤(CIK)细胞的有效策略。在这项工作中,通过摇瓶在摇床上进行动态悬浮培养,分析了其对 CIK 细胞的扩增特性和生理功能的影响,旨在优化 CIK 细胞体外扩增的培养条件。结果表明,动态培养中总细胞的扩增倍数在第 14 天达到 69.36±30.36 倍,明显高于静态培养中的 9.24±1.12 倍(P<0.05),但两种培养方法中 CD3 细胞和 CD3CD56 细胞的比例相似,导致动态培养中 CD3 细胞和 CD3CD56 细胞的扩增倍数更高。此外,两种培养方法中扩增的 CIK 细胞具有相似的生理功能。值得注意的是,动态培养中 CD25 细胞和 CD69 细胞的比例明显更高(P<0.05)。此外,动态培养中细胞的葡萄糖消耗率明显更高(P<0.05),但 Y 相似。进一步的,动态培养中的细胞具有更好的葡萄糖利用效率。综上所述,这些结果表明,动态培养增强了细胞的激活程度,从而加速了葡萄糖的消耗率,这促进了细胞的扩增和提高了细胞对葡萄糖的利用效率。

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