不同孔径微孔滤膜对肿瘤细胞滤过的影响

摘要

目的 探讨不同孔径微孔滤膜对不同直径大小的肿瘤细胞的滤过作用及其对过滤后细胞活性的影响.方法 选择不同孔径(1、3、5、8、10 μm)的聚碳酸酯微孔滤膜分别过滤浓度相同但细胞直径不同的Jurkat、K562和A549细胞,测定细胞通过滤膜的滤过率;光学显微镜测量3种细胞(包括甲醛固定后的K562)的直径及其经不同孔径的滤膜过滤后的细胞直径;台盼蓝染色法检测经滤膜过滤后的K562细胞活性,对细胞进行再培养,并用生长曲线分析其细胞增殖能力.结果 Jurkat、K562和A549细胞均不能通过1μm孔径的滤膜,通过3、5、8、10 μm孔径滤膜的滤过率均依次升高.经3μm孔径滤膜滤过的K562细胞存活率为92.0％,且滤过后的细胞再培养后的细胞增殖能力仍较强.经甲醛固定的K562细胞的滤过率明显降低,且细胞平均直径无明显变化.结论 活细胞能穿过孔径小于其直径的滤膜;穿过滤膜的细胞仍保持活性增殖,但甲醛固定可明显减少细胞通过滤膜.%Objective To investigate the filtration roles of microporous membranes with different pore sizes in the tumor cells with different diameters,and effects on the filtered cells.Methods Three kinds of tumor cells with different cell diameters and same concentrations,including Jurkat,K562 and A549,were filtered by the polycarbonate microporous membranes with different pore sizes such as 1,3,5,8 and 10 μm,respectively,and their filtration rates were determined.The diameters of three kinds of tumor cells before and after filtration,and the fixed K562 cells with formaldehyde,were measured by an optical microscope.The activity of the filtered K562 cells were detected by the trypan blue staining.After the filtered K562 cells were re-cultured,their proliferation activity was analyzed by the growth curve.Results Jurkat,K562 and A549 cells couldn't pass the filter membrane with 1 μm of pore size.The filtration rates of three kinds of tumor cells passing the fliter membranes with 3 μm,5 μm,8 μm and 10 μm of pore sizes increased in turn.The survival rate of K562 cells filtered by 3 μm of pore size of membrane was 92.0％,and the proliferation acticity of re-cultured K562 cells was still strong.The filtration rate of the fixed K562 cells with formaldehyde was significantly decreased,and the average diameter of the filtered cells had no obvious change.Conclusion The living cells are able to pass the membranes with the pore sizes less than their diameters.The living cells passed the filter membranes may still maintain their growth and proliferation activity.However,the fixation of formaldehyde may significantly reduce the number of cells passed the membrane.