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目的:研究-80℃冰箱直接冻存对细胞因子诱导的杀伤细胞(CIK)杀伤活性的影响。方法采用非程序降温方法-80℃冰箱冻存CIK,于冻存后6、12、18周复苏,通过LDH释放法分别测定其对K562细胞的杀伤活性,与新鲜未经冻存的CIK的杀伤活性进行比较。结果:未冻存的CIK在培养至第11天时,有较多贴壁成团细胞,而冻存后复苏培养的CIK贴壁成团细胞较少。比较各组细胞杀伤活性,冻存6、12周后复苏的CIK与未冻存的CIK,对K562细胞的杀伤活性无显著性差异(P>0.05),冻存18周后复苏的CIK的杀伤活性与未冻存的CIK对K562细胞的杀伤活性有显著性差异(P<0.05)。结论:采用-80℃冰箱直接冻存CIK,在12周内复苏应用于临床治疗较好。
Objective: To investigate the effect of direct cryopreservation at -80 ℃ on cytokine-induced killer (CIK) cytotoxicity. Methods CIKs were cryopreserved in a freezer at -80 ℃ and resuscitated at 6, 12 and 18 weeks after cryopreservation. The killing activity of K562 cells on K562 cells was determined by LDH release assay, Activity comparison. Results: The non-cryopreserved CIK had more adherent mesenchymal cells on the 11th day after culturing, but less CIK adherent mesenchymal cells after cryopreservation. Compared with the cytotoxic activity of each group, there was no significant difference in killing activity of K562 cells (P> 0.05) between the CIK recovered after 6 and 12 weeks of cryopreservation and the non-frozen CIK, and the killing of CIK recovered after 18 weeks cryopreservation The cytotoxic activity of CIK against K562 cells was significantly different between active and non-frozen CIK cells (P <0.05). Conclusion: The direct use of -80 ℃ refrigerator cryogenic CIK, 12 weeks of recovery for clinical treatment is better.