血清来源对细胞因子诱导的杀伤细胞扩增和功能的影响。
The effect of serum origin on cytokines induced killer cell expansion and function.
机构信息
Gene Therapy Research Center, Digestive Disease Research Institute, Tehran University of Medical Sciences, Shariati Hospital, North Kargar Ave, Tehran, 14117, Iran.
Department of Hematology, School of Allied Medical Sciences, Tehran University of Medical Sciences, Tehran, Iran.
出版信息
BMC Immunol. 2023 Sep 1;24(1):28. doi: 10.1186/s12865-023-00562-3.
BACKGROUND
Cytokine-induced killer (CIK) cells have shown promising results in adoptive immunotherapy. However, serum may play a determining role in the large-scale expansion of these cells for clinical applications. According to Good Manufacturing Practice (GMP) guidelines to reduce the use of animal products in cell-based therapies; therefore, this study sought to investigate the impact of serum origin and the reduced serum concentration on the pattern of cell expansion and function.
METHODS
Peripheral blood mononuclear cells (PBMCs) isolated from a healthy donor were expanded based on the CIK cell expansion protocol. The cell culture medium was supplemented with three types of sera comprising fetal bovine serum (FBS), human serum (HS), or human-derived platelet lysate (hPL) at different concentrations (10%, 5%, and 2.5%). The proliferation kinetics for each group were investigated for 30 days of cell culture.
RESULTS
Cell proliferation in 10% concentration of all sera (hPL, FBS, HS) was higher than their lower concentrations. Moreover, hPL was significantly associated with higher expansion rates than FBS and HS in all three concentrations. Furthermore, cells cultured in hPL showed higher viability, cytotoxicity effect, and CIK CD markers expression.
CONCLUSION
hPL at a concentration of 10% showed the best effect on CIK cell proliferation and function.
背景
细胞因子诱导的杀伤（CIK）细胞在过继免疫治疗中显示出良好的效果。然而，血清可能在这些细胞的大规模扩增中起决定性作用，以满足临床应用的需求。根据良好生产规范（GMP）指南，为了减少细胞治疗中动物产品的使用；因此，本研究旨在探讨血清来源和降低血清浓度对细胞扩增和功能模式的影响。
方法
从健康供体中分离外周血单核细胞（PBMC），并根据 CIK 细胞扩增方案进行扩增。细胞培养培养基中添加三种血清，包括胎牛血清（FBS）、人血清（HS）或人源血小板裂解液（hPL），浓度分别为 10%、5%和 2.5%。研究了每组细胞在 30 天的细胞培养过程中的增殖动力学。
结果
所有血清（hPL、FBS、HS）的 10%浓度下的细胞增殖率均高于较低浓度。此外，在所有三种浓度下，hPL 与更高的扩增率显著相关，优于 FBS 和 HS。此外，在 hPL 中培养的细胞表现出更高的存活率、细胞毒性作用和 CIK CD 标志物表达。
结论
浓度为 10%的 hPL 对 CIK 细胞的增殖和功能具有最佳效果。
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