{"title":"[<i>In Vitro</i> Amplification of NK Cells from Feeder Layer Cells Expressing IL-21].","authors":"Zhen-Zhao Xu, Xue-Hua Zhang, Ling-Ping Zhao, Gao-Hua Li, Tian-Tian Cui, Xiao-Ling Wang, Xuan Li, Ru-Ge Zang, Wen Yue, Ya-Nan Wang, Guo-Xin Li, Jia-Fei Xi","doi":"10.19746/j.cnki.issn.1009-2137.2024.05.043","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To investigate the effect of feeder layer cells expressing interleukin (IL)-21 on the amplification of NK cells <i>In Vitro</i> .</p><p><strong>Methods: </strong>The K562 cell line with IL-21 expression on its membrane was constructed by electroporation, and co-cultured with NK cells after inactivation. The proliferation of NK cells was observed. The killing function of the amplified NK cells <i>In Vitro</i> was evaluated by the lactate dehydrogenase (LDH) and interferon-γ (IFN-γ) release assay. A colorectal cancer xenograft model in NOD/SCID mice was established, and a blank control group, a NK cell group and an amplified NK cell group were set up to detect the tumor killing effect of amplified NK cells <i>in vivo</i>.</p><p><strong>Results: </strong>K562 cells expressing IL-21 on the membrane were successfully constructed by electroporation. After co-culturing with K562 cells expressing IL-21 on the membrane for 17 days, the NK cells increased to 700 times, which showed an enhanced amplification ability compared with control group (<i>P</i> < 0.001). In the tumor cell killing experiment <i>In Vitro</i> , there was no significant difference in the killing activity on tumor cells between NK cells and amplified NK cells, and there was also no significant difference in mice <i>in vivo</i>.</p><p><strong>Conclusion: </strong>K562 cells expressing IL-21 on the membrane can significantly increase the amplification ability of NK cells <i>In Vitro</i> , but do not affect the killing function of NK cells <i>In Vitro</i> and <i>in vivo</i>. It can be used for the subsequent large-scale production of NK cells <i>In Vitro</i> .</p>","PeriodicalId":35777,"journal":{"name":"中国实验血液学杂志","volume":"32 5","pages":"1578-1584"},"PeriodicalIF":0.0000,"publicationDate":"2024-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"中国实验血液学杂志","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.19746/j.cnki.issn.1009-2137.2024.05.043","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q4","JCRName":"Medicine","Score":null,"Total":0}
引用次数: 0
Abstract
Objective: To investigate the effect of feeder layer cells expressing interleukin (IL)-21 on the amplification of NK cells In Vitro .
Methods: The K562 cell line with IL-21 expression on its membrane was constructed by electroporation, and co-cultured with NK cells after inactivation. The proliferation of NK cells was observed. The killing function of the amplified NK cells In Vitro was evaluated by the lactate dehydrogenase (LDH) and interferon-γ (IFN-γ) release assay. A colorectal cancer xenograft model in NOD/SCID mice was established, and a blank control group, a NK cell group and an amplified NK cell group were set up to detect the tumor killing effect of amplified NK cells in vivo.
Results: K562 cells expressing IL-21 on the membrane were successfully constructed by electroporation. After co-culturing with K562 cells expressing IL-21 on the membrane for 17 days, the NK cells increased to 700 times, which showed an enhanced amplification ability compared with control group (P < 0.001). In the tumor cell killing experiment In Vitro , there was no significant difference in the killing activity on tumor cells between NK cells and amplified NK cells, and there was also no significant difference in mice in vivo.
Conclusion: K562 cells expressing IL-21 on the membrane can significantly increase the amplification ability of NK cells In Vitro , but do not affect the killing function of NK cells In Vitro and in vivo. It can be used for the subsequent large-scale production of NK cells In Vitro .
目的研究表达白细胞介素(IL)-21 的饲养层细胞对体外 NK 细胞扩增的影响:用电穿孔法构建膜上表达 IL-21 的 K562 细胞系,灭活后与 NK 细胞共培养。观察了 NK 细胞的增殖情况。通过乳酸脱氢酶(LDH)和干扰素-γ(IFN-γ)释放试验评估了体外扩增的 NK 细胞的杀伤功能。建立了 NOD/SCID 小鼠结直肠癌异种移植模型,并设置了空白对照组、NK 细胞组和扩增 NK 细胞组,以检测扩增 NK 细胞在体内的肿瘤杀伤作用:结果:通过电穿孔成功构建了膜上表达IL-21的K562细胞。结果:电穿孔成功构建了膜上表达IL-21的K562细胞,与膜上表达IL-21的K562细胞共培养17天后,NK细胞增至700倍,与对照组相比,扩增能力增强(P < 0.001)。在体外肿瘤细胞杀伤实验中,NK细胞和扩增的NK细胞对肿瘤细胞的杀伤活性无显著差异,在小鼠体内也无显著差异:结论:膜上表达 IL-21 的 K562 细胞能显著提高 NK 细胞的体外扩增能力,但不影响 NK 细胞在体外和体内的杀伤功能。它可用于随后大规模生产体外 NK 细胞。