摘要
目的探讨脐带血、健康人和肝癌患者外周血来源的细胞因子诱导的杀伤细胞(CIK)在体外的增殖能力,及其对肝癌SMMC7721细胞的杀伤效应。方法无菌采集脐带血、健康人和肝癌患者的外周血,用淋巴细胞分离液分离单个核细胞,加入相应的细胞因子体外诱导CIK,细胞计数法检测CIK的增殖能力,流式细胞术检测细胞的表型,四甲基偶氮唑蓝法检测诱导的CIK对SMMC7721细胞的杀伤效应。结果诱导14 d后,脐带血、健康人和肝癌患者外周血来源的CIK细胞增殖倍数分别为153.8±13.1、137.6±10.3和95.3±8.5,经两两比较差异均有统计学意义(P<0.05)。诱导培养14 d后,脐带血、健康人和肝癌患者外周血来源单个核细胞的CD3^+CD56^+表型细胞所占百分比分别为(45.3±4.6)%、(36.2±2.8)%、(21.9±2.7)%,CD3^+CD56^+表型细胞所占百分比较诱导前有显著提高,差异均有统计学意义(P<0.05);脐带血来源的CIK中CD3^+CD56^+表型细胞所占比例显著高于健康人和肝癌患者,差异均有统计学意义(P<0.05)。经14 d的诱导培养后,3种来源的CIK对肝癌细胞SMMC7721的杀伤活性经两两比较差异均有统计学意义(P<0.05),脐带血的杀伤活性最强,健康人的杀伤活性次之,肝癌患者的杀伤活性最弱。在效靶比10:1时杀伤活性强于效靶比5:1(P<0.05)。结论脐带血来源的CIK体外增殖快,对肝癌细胞SMMC7721的杀伤活性强,是肿瘤细胞生物治疗的最佳选择。
Objective To explore the reproductive activity in vitro and killing effects to hepatoma cell SMMC7721 of cytokine-induced killer cell（CIK） from cord blood,peripheral blood of health people and liver cancer patients.Methods The cord blood and peripheral blood of health people and patients with hepatoma were collected under asepsis condition and the mononuclear cells were separated by lydroxypropylmethyl cellulose.CIK were induced by adding a variety of cytokine in different time.The reproductive activity of CIK were measured by cell counting.Flow cytometry was used to detect the cell phenotype of CIK.The methyl thiazolyl tetrazolium was used to detect the killing effect of CIK to SMMC7721 cells.Results After 14 days of induction,the proliferation multiple of CIK from cord blood,peripheral blood of health people and liver cancer patients was 153.813.1,137.6±10.3 and 95.3±8.5 respectively,the difference of them was statistic significance（P0.05）.After 14 days of induction and cultivation,the percentage of mononuclear cells with CD3~＋ CD56~＋ phenotype from cord blood,peripheral blood of health people and liver cancer patients was（45.3 ±4.6）%,（36.2±2.8）%,（21.9±2.7）%respectively, and it was significantly higher than before induction（P0.05）;the percentage of mononuclear cells with CD3~＋ CD56~＋ phenotype from cord blood was significantly higher than that from peripheral blood of health people and liver cancer patients（P0.05）.After 14 days induction and cultivation,the differences of the killing effect of CIK from three source to hepatoma cell SMMC7721 cells were statistic significance（P0.05）.The killing activity of cord blood was strongest,health people followed, liver cancer patients was weakest.The killing activity at effect-target ratios 10：1 was stronger than at 5：1（P 0.05）.Conclusion CIK derived from cord blood possess high speed of proliferation in vitro and strong killing effect to hepatoma cell SMMC7721,thus being the best choice for tumor biologic therapy.
出处
《新乡医学院学报》
CAS
2013年第10期801-803,共3页
Journal of Xinxiang Medical University
基金
新乡市科技局科技攻关项目(编号:09S052)
