人外周血与脐带血来源自然杀伤细胞大体系扩增培养比较

Large-scale amplification culture of the natural killer cells from human peripheral blood and cord blood

目的探讨人外周血、脐带血来源的细胞因子诱导的自然杀伤(NK)细胞在体外大体系扩增培养的增殖能力及细胞表型分析。方法无菌采集健康志愿者外周血、孕产妇脐带血,用淋巴细胞分离液分离单个核细胞,加入白细胞介素-2(IL-2)体外诱导NK细胞,细胞计数法检测比较其增殖能力,流式细胞术检测诱导后NK细胞的表型。结果加入IL-2诱导体外培养15 d后,外周血、脐带血来源的NK细胞扩增倍数分别为187.87±7.57和165.80±9.24,外周血来源的NK细胞扩增倍数高于脐带血来源的(P<0.05)。培养前,流式细胞术检测外周血、脐带血不同来源单个核细胞中CD3^-CD16^+CD56^+表型的NK细胞所占比例分别为(18.37±3.16)%和(13.29±2.41)%,差异无统计学意义(P>0.05);诱导培养15 d后,外周血、脐带血不同来源单个核细胞中CD3^-CD16^+CD56^+表型的NK细胞所占比例分别为(91.32±5.38)%和(84.26±4.11)%,较诱导前均显著升高(P<0.05);外周血来源CD3^-CD16^+CD56^+表型的NK细胞所占比例稍高于脐带血来源,但差异无统计学意义(P>0.05)。结论经细胞因子IL-2诱导后,外周血和脐血来源诱导的NK细胞的增殖能力、CD3^-CD16^+CD56^+表型细胞所占比例均有较大提高。

Objective To investigate the proliferation ability of the natural killer（ NK） cells which from human peripheral blood and cord blood induced by the cytokine and their phenotype. Methods Aseptic collection of peripheral blood from healthy volunteer and cord blood from parturient. The mononuclear cells were isolated by the lydroxypropylmethyl cellulose;then added in NK cells which induced by interleukin-2（ IL-2） in vitro. The ability of proliferation of NK cells was detected by cytometry and the phenotype of the NK cells was detected by flow cytometer. Results The proliferation time of NK cells which derived from peripheral blood and cord blood was 187. 87 ± 7. 57 and 165. 80 ± 9. 24 respectively after being cultured for 15 days in vitro; the proliferation time of NK cells which derived from peripheral blood was significantly higher than that derived from the cord blood（ P 〈 0. 05）. There was no statistic difference in the ratio of CD3-CD16＋CD56＋phenotype NK cells in mononuclearcell from peripheral blood[（ 18. 37 ± 3. 16） %]and cord blood[（ 13. 29 ± 2. 41） %]before culturing. After culturing for 15 days,the ratio of CD3-CD16＋CD56＋phenotype NK cells in mononuclearcell from peripheral blood and cord blood was（ 91. 32 ±5. 38） % and（ 84. 26 ± 4. 11） % respectively,which was significantly higher than before culturing（ P 〈 0. 05）. The ratio of CD3-CD16＋CD56＋phenotype NK cells in mononuclearcell from peripheral blood was more than that from cord blood,but there was no statistic difference（ P 〉 0. 05）. Conclusion The proliferation activity of NK cells and the ratio of CD3-CD16＋CD56＋phenotype NK cells which derived from peripheral blood and cord blood is increased after being induced by IL-2.