不同保存时间去白红细胞上清对急性T淋巴细胞白血病Jurkat细胞增殖和凋亡的影响

Effect of supernatants of leukoreduced red blood cells of different storage time on proliferation and apoptosis of acute T-lymphocyte leukemia Jurkat cells in vitro

目的采用体外实验研究不同保存时间去白细胞(去白)红细胞上清对人T淋巴细胞株Jurkat细胞增殖及凋亡的影响。方法设含10%灭活胎牛血清的1640培养液为空白组,单纯Jurkat细胞培养为对照组,保存7 d去白细胞悬浮红细胞上清与Jurkat细胞共培养为实验A组,保存30 d去白细胞悬浮红细胞上清与Jurkat细胞共培养为实验B组。采用细胞计数试剂盒(CCK-8)法检测各组培养24、48 h后对Jurkat细胞增殖的影响。流式细胞术检测红细胞上清作用于Jurkat细胞48 h后对细胞凋亡的影响。结果在开始培养时各组生长抑制率无明显差异(P>0.05);培养24、48 h后,实验B组生长抑制率明显低于对照组[(5.72±0.76)%vs(8.42±0.73)%;(4.71±0.54)%vs(9.16±0.86)%;P均<0.05]。实验A组与对照组生长抑制率相当(P>0.05)。在培养开始时各组Jurkat细胞凋亡率无明显差异(P>0.05);培养48 h后实验B组凋亡率明显低于对照组[(6.71±1.14)%vs(10.12±1.67)%,P<0.05],而实验A组与对照组细胞凋亡率相当(P>0.05)。结论新鲜去白红细胞上清对Jurkat细胞增殖和凋亡无明显影响,陈旧的去白红细胞上清对Jurkat细胞的增殖有促进作用,对其凋亡有抑制作用,且这种作用存在一定的时间依赖性。

Objective To investigate the effect of supernatants of red blood cells of different storage time on proliferation and apoptosis of acute T-lymphocyte leukemia cell strain Jurkat cells in vitro. Methods The 1640 medium containing10% inactivated fetal calf serum was served as blank group,The Jurkat cells of pure culture were served as control group.The supernate of leukoreduced suspended erythrocytes stored for 7 days co-cultured with Jurkat cells was served as experiment group A. The supernate of leukoreduced suspended erythrocytes stored for 30 days co-cultured with Jurkat cells was served as experiment group B. The cell counting kit-8（CCK-8） was used to detect the effects on proliferation of Jurkat cells in each group 24 and 48 hours aftrcultire. Flow cytometry was used to detect the effect on apoptosis of Jurkat cells after supernate of leukoreduced erythrocytes acting on Jurkat cells for 48 hours. Results There were no significant differences in cell growth inhibition rates for all groups at the beginning of culture（P〉0. 05）. After cultures of 24 and 48 hours,cell growth inhibition rates in experiment group B were significantly lower than those in control group [（5. 72 ± 0. 76） % vs（8. 42 ± 0. 73） %;（4. 71 ± 0. 54） % vs（9. 16 ± 0. 86） %; all P〈0. 05],and this showed that the erythrocytes supernate stored for 30 days had a certain promoting effect on the proliferation of Jurkat cells with a certain time dependence. There were no significant differences in cell growth inhibition rates after cultures of 24 and 48 hours between experiment group A and control group（all P〉0. 05）. There was no significant difference in apoptosis rates of Jurkat cells at the beginning of culture between experiment group A and experiment group B（P〉0. 05）,and the cell apoptosis rates of Jurkat cells in experiment group B was significantly lower than that in control group after cultures of 48 hours [（6. 71 ± 1. 14） % vs（9. 16 ± 0. 86） %,P〈0. 05],while there was no significant difference between experiment group and control group（P〉0. 05）. This showed that the erythrocytes supernate stored for 30 days had inhibiting effect on cell apoptosis. Conclusion The fresh erythrocytes supernate has no obvious effect on the proliferation and apoptosis of Jurkat cells,while the old erythrocyte supernate might have the effect of promoting proliferation and inhibiting apoptosis on Jurkat cells.