【摘要】 目的:脐血处理的关键问题是提高干细胞的回收率及实现处理过程的标准化和可重复化,实验对此进行探讨,比较干细胞分离仪与传统羟乙基淀粉手工法分离脐血的效果。方法:实验于2006-12/2007-05在广州医学院附属市一人民医院完成。①脐血来源:39份脐血采自广州医学院附属市一人命医院妇产科健康顺产新生儿脐带,产妇均知情同意。随机数字表法分为仪器分离组17份、手工分离组22份。②实验方法:仪器分离组收集脐血称质量,计算体积,在开始处理前20min缓慢加入相当于20%脐血体积的60g/L羟乙基淀粉。仪器分离组按仪器要求自动分离,分离终体积20mL。手工分离组50g离心5min,压浆板压出全部血浆以及18mL红细胞移至无菌空袋,500g离心13min,自动压浆板压出血浆,保留20mL终体积样本。③实验评估:采用全自动计数仪进行检测有核细胞(白细胞)、红细胞数量。流式细胞仪分析CD34+含量。结果:采用干细胞分离仪处理浓缩脐血,有核细胞回收率为(89.7±3.4)%,CD34+细胞回收率为(98.8±5.1)%,红细胞去除率为(55.2±16.7)%,均比手工分离组分离效果好,差异有显著性意义(P<0.05或0.01)。同时,仪器分离组有核细胞回收率、CD34+细胞回收率的标准差均明显低于手工分离组(3.4vs.15.3;5.1vs.10.3)。结论:相比传统的羟乙基淀粉手工分离法,干细胞分离仪脐血分离浓缩效果理想,且结果标准误小,数据稳定。更多还原

【Abstract】 AIM:The key problem of processing umbilical cord blood is to elevate the recovery rate of stem cells and implement the standardization and repetition in the process.This experiment is designed to investigate and compare the umbilical cord blood separation effect by a new full-automatic cell separation system versus the manual hydroxyethyl starch(HES)separation method.METHODS:The experiment was carried out at Guangzhou First People’s Hospital,Guangzhou Medical College from December 2006 to May 2007.①All 39 cord blood samples were collected at the end of full-time deliveries from healthy donors at Guangzhou First People’s Hospital,Guangzhou Medical College after their agreements.There were two groups,full-automatic cell separation system group(n =17)and manual separation group(n =22).②60 g/L HES was slowly added into all samples at 20% cord blood volume,20 minutes later,the separation started after weighting cord blood mass and calculating the volume.Full-automatic cell separation system group was carried according to its handbook,and its last volume was 20 mL.The manual separation group was centrifugated at 50 g for 5 minutes,all plasm and 18 mL red cell was separated to a new bag,another 500 g for 13 minutes centrifugation,20 mL volume of cells was collected.③Numbers of nucleated and red cells were detected with Hematology Analyzer and content of CD34+ cells was mesured with flow cytometer.RESULTS:Recovery rate of nucleated cells was(89.7±3.4)%,recovery rate of CD34+ cells was(98.8±5.1)% and the removal rate of red cells was(55.2±16.7)% in the full-automatic cell separation system group.These showed better isolation effect than the manual separation group(P < 0.05 or 0.01).Simultaneously,standard deviation of recovery rates of nucleated cells and CD34+ cells was significnatly lower in the full-automatic cell separation system group than the manual separation group(3.4 vs.15.3;5.1 vs.10.3).CONCLUSION:The full-automatic cell separation system is an ideal device for processing of umbilical cord blood compared with the tranditional manual separation.It shows small standard deviation and exact data.更多还原