白细胞滤器去除血液肿瘤细胞的实验研究  

Removal oftumor cells from blood by leukocyte filter

作　　者：尚玉 邵小宝[3] 周琳 严枫 朱培元[3] SHANG Yu;SHAO Xiaobao;ZHOU£in;YAN Feng;ZHU Peiyuan(Department of Clinical Laboratory,Affiliated Cancer Hospital of Nanjing Medical University,Jiangsu Cancer Hospital,Nanjing 210009,China;Department of Clinical Laboratory,Nanjing Intergrated Traditional Chinese and Western Medicine Hospital Affiliated to Nanjing University of Chinese Medicine;Department of Transfusion Medicine,Nanjing Hospital of Chinese Medicine Affiliated to Nanjing University of Chinese Medicine)

机构地区：[1]南京医科大学附属肿瘤医院江苏省肿瘤医院检验科,江苏南京210009 [2]南京中医药大学附属南京市中西医结合医院检验科,江苏南京210014 [3]南京中医药大学附属南京中医院输血科,江苏南京210022

出　　处：《中国输血杂志》2021年第5期452-455,共4页Chinese Journal of Blood Transfusion

基　　金：南京市科技计划项目(201715036)。

摘　　要：目的了解白细胞滤器去除血液中肿瘤细胞的效果及其用于肿瘤患者术中回收式自体输血的可行性。方法将体外培养的人结肠癌细胞HCT116加入200 mL人全血制备的去白细胞悬浮红细胞(简称去白悬红)中,模拟肿瘤患者术中回收的自体血液。分别采用常规(A组,n=6),滤膜孔径较小(B组,n=6)和滤膜较厚且孔径较小(C组,n=6)3种白细胞滤器(简称滤器)过滤;利用抗-CD45-PE、抗-CD326(EpCAM)-FITC流式细胞术检测3组中的CD45-EpCAM+肿瘤细胞数量,以Flow Count荧光计数微球做绝对定量计数,采用细胞培养和台盼蓝染色镜检检测肿瘤细胞活性,全自动生化仪速率法检测血液乳酸脱氢酶(LDH)、离子选择性电极间接法测K^(+)浓度,称重法计算血液回收率。结果流式细胞计数:A、B、C 3种(组)滤器滤除前血液中的肿瘤细胞数(×10^(7)/mL)分别为1.52±0.48、1.48±0.55、1.44±0.46,滤除后的肿瘤细胞数(×10^(4)/mL)分别为2.73±1.74、2.96±1.85、3.08±2.33(P<0.01)。A、B、C 3组过滤后的血液培养7 d均未见存活肿瘤细胞,过滤前后3个组之间K^(+)、LDH含量相近(P>0.05),3组的血液回收率分别为85.22%、84.97%、82.86%。结论常规使用的滤器可明显降低循环肿瘤细胞的数量,用于肿瘤患者术中自体血回收输注具有一定可行性。Objective To investigate the effect of leukocyte filter in removal of tumor cells in blood and the viability of using leukocyte filter in intraoperative cell salvage in tumor patients.Methods The leucocyte-depleted suspended RBCs prepared from 200 mL human whole blood were spiked with HCT116 cells cultured in vitro to simulate the autologous blood recovered from tumor patients during operation.Conventional filter(group A,n=6),filter with small pore size(group B,n=6) and filter with thicker membrane and small pore size(group C,n=6)were used.The CD45-EpCAM+tumor cells were detected by anti-CD45-PE and anti-CD326(EpCAM)-FITC flow cytometry,and the absolute count of tumor cells were calculated using Flow Count fluorescence microspheres.The activity of tumor cells was detected by cell culture and trypan blue staining.Furthermore,blood biochemical indexes(LDH,K^(+))and red cell recovery rate were detected.Results Tumor cell counts before and after the application of leukocyte filter A,B,C were(1.52±0.48)×10^(7)/mL vs(2.73±1.74)×10^(4)/mL,(1.48±0.55)×10^(7)/ml vs(2.96±1.85)×10^(4)/mL and(1.44±0.46)×10^(7)/mL vs(3.08±2.33)×10^(4)/mL,respectively(P>0.05),which was significantly reduced after filtration(P<0.01).No viable cells were found in the filtered blood cultured for 7 days.There were no significant differences in K^(+)and LDH value before and after filtration among the three groups(P>0.05).The blood recovery rates of group A,B,C were 85.22%,84.97% and 82.86%,respectively.Conclusion The conventional filter can significantly reduce the number of circulating tumor cells,and it is feasible for intraoperative cell salvage in tumor patients.

关 键 词：术中自体血回收 肿瘤细胞去除 白细胞滤器 红细胞回收率 流式细胞术 

分 类 号：R457.12[医药卫生—治疗学] R730.1[医药卫生—临床医学]