观察不同保存期血液诱导的CIK细胞功能及对卵巢癌SKOV3ip细胞的影响  

Immuno-Function of Cytokine-Induced Killer Cells in Different Retention Period and Its Lethal Effect to SKOV3ip Cells

作　　者：柯雪[1] 杨琨[2] 董武[2] 

机构地区：[1]大连医科大学研究生院,辽宁大连116044 [2]辽宁省人民医院,辽宁沈阳110016

出　　处：《实用妇产科杂志》2014年第2期137-140,共4页Journal of Practical Obstetrics and Gynecology

摘　　要：目的：了解不同保存期机采血诱导的细胞因子诱导的杀伤细胞（CIK）激活率、体外增殖情况及对卵巢癌细胞株SKOV3ip的体外细胞毒性。方法：抽取10例卵巢癌患者机采血各15ml（共10份），复方枸橼酸钠抗凝，置4℃保存，于采血后的第0～9天每日每份均取1rnl血液，常规分离单个核细胞（PBMC），并计数，加入细胞因子干扰素（IFN）、白细胞介素-2（IL-2）、抗-CD3单抗（CD3McAb）和IL-1，体外诱导CIK。流式细胞术检测其细胞激活率、对比不同保存期机采血诱导的CIK的增殖能力、M1Tr比色法检测其对卵巢癌SKOV3ip细胞的杀伤力。结果：机采血PBMC计数第6天开始死亡率显著上升；保存第7天时CIK激活率下降显著，保存第9天时PBMC全部不能激活诱导为CIK；保存第7天CIK对SKOV3ip杀伤能力明显下降，第9天已无杀伤力。随着血液保存时间的延长，CIK增殖能力逐渐减低，至第8天不能再增殖。但保存4天内的血液这几项指标变化不大（P〉0．05）。结论：随着保存时间的延长，机采血诱导培养CIK的激活率、增殖能力及杀伤力均呈递减趋势。制备CIK的机采血在保存4天内诱导的CIK功能变化不大。Objective：To investigate the cell activation rate, proliferation capacity in vitro and the cytotoxic- ity on SKOV3ip ovarian cancer cell lines of CIK in different retention period. Methods： Blood samplings from 10 normal human （each 15 ml） were extracted,anticoagulated with compound folic acid sodium and pre- served at 4℃. PBMC were separated and counted from 1 ml blood each day and cell factors IFN, IL-2, CD3McAb and IL-1 were added for induce PBMC to CIK cells in vitro. Flow cytometry was used to test the cell activation rate. Proliferation capacity of CIK cell in different retention period were compared and cytotoxici- ty on SKOV3ip ovarian cancer cell lines were evaluated by MTT. Results： Mortality rate of PBMC began to rise significantly on the sixth day;its activation percentage dropped from the seventh day,and CIK cell could not be activated from PBMC from the ninth day;lethal effect dropped down from the seventh day, and CIK cell had no cytotoxicity from the ninth day;as the prolongation of storage period,the cell proliferation ability gradually declined,CIK could not be induced on the eighth day; however,the aforementioned indicators did not change significantly during the first four days （ P 〉 0.05）. Conclusions： Along with the prolongation of storage period,cell activation rate, proliferation capacity in vitro and the cytotoxicity of CIK cells gradually de- creased. The immuno-function of CIK cell has no change in four-day storage period.

关 键 词：细胞因子诱导的杀伤细胞 细胞激活率 杀伤力 增殖能力 SKOV3ip卵巢癌细胞 

分 类 号：R737.31[医药卫生—肿瘤]