A-NK细胞培养和活化方法的改进

The improvement in culture and activation of A-NK cells

目的:比较A-NK细胞在完全培养基(CM)和无血清培养基(AIMV)中体外扩增及杀伤肿瘤细胞作用,同时探讨IL-12对于A-NK/IL-2治疗的辅助作用及其杀伤的形态学观察。方法:用MTT法比较不同培养条件下的A-NK细胞体外增殖能力,并测定细胞体外杀伤肿瘤细胞的活性。通过做扫描和透射电镜,对A-NK细胞杀伤的肿瘤细胞进行形态学观察。结果:不同培养条件下的A-NK细胞均可在短期内大量扩增(P<0.05),且均有很强的杀伤肿瘤的活性(P<0.05)。被A-NK细胞杀伤的肿瘤细胞的死亡形式是溶解坏死(necrosis)和凋亡(apoptosis)。结论:AIMV可替代CM用于A-NK细胞的培养。联合应用IL-12和IL-2激活A-NK细胞优于单独使用IL-2,可减轻高剂量IL-2带来的副作用。此研究为A-NK细胞的实验室研究和将来临床应用推广奠定了新的实验基础。

Objective:To compare expansion and kill activation of A-NK cells cultured with serum-free medium AIMV and standard serum-containing medium in vitro, and also detest the assist effect of EL-12 on A-NK/IL-2 treatment and the morphology character of killed cells through electroscope.Methods:Proliferation ability of the cells cultured in the two different media was compared by MTT method in vitro. The morphology of the killed tumor target cells by adherent natural killer cells (A-NK) was observed through electroscope. Results:All of A-NK cells cultured in deferent condition could rapidly proliferate(P<0.05) and kill target cells(P<0.05),Death patterns of tumor target cells were necrosis and apoptosis.Conclusion:The serum-free medium AIMV could replace standard serum-containing medium for culturing A-NK cells in vitro, A-NK cells activated by both IL-2 and IL-12 was superiority compare the cells activated by IL-2 only,and could reduce side effects by injecting of high dose EL-2. The studies provided a new experimental basis for laboratory and apply of clinical treatment of A-NK cell in future.