无菌塑料袋分离单个核细胞

Separation of mononuclear cells using bacteria-free plastic bags

目的:研究适合用于人体的利用无菌塑料袋分离单个核细胞(MNC)的方法。方法:随机抽查20份样本,单份采集量65～400ml,分为2组,15～58岁为第1组(n=15),59～76岁为第2组(n=5)。将Ficoll淋巴细胞分离液加入一次性无菌塑料袋内,将采集的血液或骨髓样本缓慢加入淋巴细胞分离液的液面上,离心,收集MNC层,然后洗去Ficoll淋巴细胞分离液,根据临床需要调整MNC的液体悬浮量。结果:第1组分离后MNC的细胞数为(4.90±2.69)×108,回收率为(71.96±12.43)%,CD34+细胞占MNC的比率为(2.31±0.93)%。第2组分离后MNC的细胞数为(1.11±0.74)×108,回收率为(69.98±2.23)%,CD34+细胞占MNC的平均比率为(0.98±0.04)%。结论:利用无菌塑料袋分离MNC,分离效果和MNC、CD34+回收率均能满足临床需要。

Aim: To research the method suitable for separating human mononuclear cells(MNC) by using bacteria-free plastic bags.Methods: Ficoll lymphocyte separation buffer (1.077 g/ml) was added into bacteria-free plastic bags.Twenty samples were divided into groupⅠ(aging from 15~58 years) and group Ⅱ(aging from 59~76 years).The collected blood or bone marrow sample were slowly added into the bacteria-free plastic bags containing Ficoll lymphocyte separation buffer, and then separated at 4 ℃,1 200 r/min.After equalization, the MNC layer was collected and washed for removing Ficoll lymophycyte separation buffer. Results: The average number of MNC, recovery rate of MNC, and the average ratio of CD34+/MNC for group Ⅰwere (4.90±2.69)×108 , (71.96±12.43)%, and (2.31±0.93)%, and these parameters for group Ⅱwere (1.11±0.74)×108, (69.98±2.23)%, and (0.98±0.04)%.Conclusion: By separating MNC using bacteria-free plastic bags, the separating efficiency and CD34+ recovery meet the clinical demand. Compared with the common method of using tub, it is free of bacteria and has no restriction of sample volume.