儿童外周血造血干细胞采集方法与效果评价

Evaluation of methods for collection of peripheral blood stem cells in children

目的:探讨儿童外周血造血干细胞(PBSC)的采集方法,并对采集效果进行评价。方法:患者采用化疗和G-CSF或GM-CSF动员,异体供者用G-CSF动员,待WBC数回升至>5.0×109/L,应用CS-3000 Plus、COM TEC和COBE Spectra 3种血细胞分离机进行PBSC采集,设定循环血量为总血容量(TBV)的2～3倍,对于体重<20 kg,红细胞压积(HCT)<24%或者TBV≤1 100～1 650 ml的小儿,须采用经25 Gyγ射线照射的同型少白细胞红细胞预注管路和低流速分离(全血流速一般为10～30 ml/min)。用流式细胞仪检测所采集干细胞的CD34+细胞数,并计数单个核细胞(MNC)数。分析采集细胞中CD34+细胞数与采前MNC数量及采集循环血量的关系。结果:使用CS-3000 Plus血细胞分离机成功采集10例,COBE Spectra血细胞分离机成功采集18例,COMTEC血细胞分离机成功采集3例。采集次数为1～5次,平均1.5次,获得CD34+细胞数(7.9±2.9)×106/kg,MNC数(7.4±3.1)×108/kg。相关性分析显示,所采集到的CD34+数量和采集前MNC数、采集的循环血量呈正相关(r2=0.6229,P<0.0001;r2=0.1905,P=0.0141)。结论:根据儿童患者/供者的体重、TBV、动员情况和循环血量等因素选择合适采集方法,经过1～2次单采都可以安全地采集到高浓度的、能满足临床需要的PBSC。

Objective： To evaluate the methods for collection of peripheral blood stem cells（PBSC） in children.Methods： Peripheral blood stem cells were collected from 20 child patients and 11 donors.The patients treated with chemotherapy,received G-CSF or GM-CSF and the donors received G-CSF for mobilization.When the peripheral blood（PB） leukocyte count reached to 5×109/L,the hematopoietic stem cells were collected with CS-3000 Plus,COM TEC or COBE Spectra blood cell separators from patients and donors.For children whose weight20 kg,HCT24% or TBV≤1 100-1 650 ml,blood cell separators were pre-injected with the same type RBCs irradiated by 25 Gy of gamma-ray and with low flow rate（10-30 ml/min）.The number of CD34＋ cell was detected by flow cytometry.The relationship of number of CD34＋ cell with mononuclear cell（MNC） and processed blood volume was analyzed.Results： Successful collection of the PBSCs with the CS-3000 Plus（n=10）,the COM TEC（n=3） and the COBE Spectra（n=18） was achieved in all the 31 cases with 1-5 aphereses used.Number of CD34＋ cells was（7.9±2.9）×106/kg and that of MNCs was（7.4±3.1）×108/kg.The total CD34＋ cell count was correlated with MNCs before aphaeresis and processed blood volume.Conclusion： For collection of high quality PBSCs,the appropriate methods should be chosen according to the body weight,TBV,mobilization of child patients/donors.