摘要
目的寻求一种CIK细胞的保存方法。方法将诱导成功的CIK细胞于14天时冻存，两个月之后复苏。观察细胞状态，复苏细胞从第一天起绘制细胞生长曲线，没有冻存的细胞从培养14天起绘制生长曲线，比较两者增殖率有无差别。复苏3天后做MTT，检测两组CIK细胞对肿瘤细胞的杀伤活性差异。复苏后第三天用流式细胞仪测定细胞表型，与没有冻存的细胞进行比较。结果复苏后细胞状态良好，与未冻存细胞相比增殖率有差异（P〈0．05）。未冻存细胞在效靶比为40：1、20：1、10：1时杀伤率分别为（51．1±2．0）％、（50．9±3．3）％、（36．4±3．2）％，冻存细胞在效靶比为40：1、20：1、10：1时杀伤率分别为（49．7±1．5）％、（34．7±1．3）％、（29．5±7．6）％，两者在效靶比不同时杀伤活性没有明显差异。细胞表型尤其是起主要杀伤作用的细胞群CD3^＋CD56^＋分别为（19．1±0．4）％、（19．5±0．8）％，两者没有明显差异。结论冻存后复苏对CIK细胞的增殖率有一定影响，但并不影响临床上的应用。对杀伤活性和细胞表型没有影响。CIK细胞冻存后复苏方便了临床的应用。
Objective To search a good way to store CIK ceils. Method Freeze CIK cells when they were induced for 14 days and anabiosis them after two months later. The anabiosis cells growth curve was drawn from the first day they recovered and the unfreezing cells growth curve was drawn after they induced for 14 days. The cytotoxic activities were determined by MTT and the cell phenotypes were detected by flow cytometry after anabiosis three days. Three indexes of anabiosis CIK cells were compared to the unfreezed. Results The anabiotic cells were in good condition. Compared with unfreezing CIK cells, the growth rate of freezed cells was lower, significant differences existed between the two groups （ P 〈 0.05 ）. Cytotoxic activities of CIK cells and the freezed ones were（51.1 ±2.0）% ,（50.9±3.3）% ,（36.4 ±3.2）% and（49.7 ±1.5）% ,（34.7 ±1.3）% ,（29.5 ±7.6）% according to the different effect and target cell ratios of 40： 1,20： 1 and 10： 1. No significant difference existed between the two groups. The cell phenotypes CD3 ^＋ CD56^ ＋ were （19.1 ± 0.4 ）%, （19.5 ± 0.8 ）% and there were no significant difference between the two groups. Conclusion Freezing had effect on the growth rate of CIK cells but had no influence on cytotoxie activity and cell phenotype. The method is available for clinical application.
出处
《实用肿瘤学杂志》
CAS
2009年第6期509-512,共4页
Practical Oncology Journal
基金
黑龙江省重点攻关课题基金资助（900C190401）
