1727份公共库脐带血的质量分析——附23份库存脐带血干细胞解冻质控分析  

Analysis on quality of 1 727 cases' umbilical cord blood in public cord blood bank-attached 23 cases on quality control of thawed cord blood stem cells in inventory

作　　者：王秋实[1] 马良艳[1] 张晋[1] 佟海侠[1] 王殿昌[1] 段小晶[1] 吴思梦[1] 袁哲[1] 王哲[1] 刘学勇[1] 

机构地区：[1]中国医科大学附属盛京医院输血科,辽宁沈阳110004

出　　处：《中国输血杂志》2014年第12期1317-1321,共5页Chinese Journal of Blood Transfusion

摘　　要：目的评估公共库存脐带血造血干细胞的质量及脐血库用于质控的血样是否能够正确反应实际血袋内的脐带血造血干细胞的活性和数量。方法分析2009年1月-2010年12月按照Rubinsten标准分离入库保存的1 727例合格脐带血造血干细胞,通过流式细胞术和集落培养法计算造血干细胞数量,血细胞计数仪计数细胞数量、应用气相液氮系统深低温冷冻造血干细胞。分析入库细胞总数(TNC)的影响因素,以及与CD34+总数、集落形成单位(CFU-C)总数的相关性;对冷冻时间>24个月的23份脐带血造血干细胞解冻后,分析解冻前后血袋、冷冻麦管内和冻存管内的细胞数、CFU-C产率、CD34+百分比、活细胞比率,并推算血袋内TNC、CD34+细胞数、CFU-C总数。结果1 727份脐带血平均血量为(96.62±20.41)m L,入库TNC、CD34+总数、CFU-C总数分别为(9.88±2.54)×108个、(2.76±1.85)×106个、(11.05±7.72)×105个,CD34+总数和CFU-C总数与TNC相系数均为0.49,CD34+百分比与CFC产率相关系数为0.80。23份解冻脐带血冷冻前后血袋、冷冻管和冷冻麦管内WBC(×109/L)分别为28.03±8.01、22.55±6.56、20.43±5.23,TNC(×108)为10.74±3.33、58±2.90、7.78±2.28、7.97±2.73、台盼兰拒染率(%)99.0±0、83.04±6.21、78.9±6.56、80.43±6.01(P<0.01);冷冻前和解冻后血袋、冷冻管,冷冻麦管的CFU-C产率(/105)分别为159.9±45.8、149.2±37.4、91.6±35.8、155.0±41.2,冷冻管内CFU-C产率明显下降(P<0.05);冷冻前和解冻后血袋、冷冻管,冷冻麦管的CFU-C总数(×105)分别为159.9±45.8、149.2±37.4、91.6±35.8、155.0±41.2,解冻后冷冻管内CFU-C总数下降明显(P<0.05);血袋、冷冻管和麦管内的小细胞数量(×108)分别为3.92±1.40、3.95±1.77、3.86±1.46、4.05±1.52,CD34+总数(×106)为3.23±1.84、3.02±1.88、3.12±1.85、3.62±3.07。结论公共库入库的1 727份脐带血符合国际通用标准;解冻后检测分析发现,脐带血冷冻后丢失的细胞以成熟的粒细�Objectives To assess the quality of cord blood hematopoietic stem cells made in a public inventory, as well as to assess whether the blood samples under quality control can accurately reflect the activity and the amount of umbilical cord blood stem ceils in the cryopreserved bag. Methods From January 2009 to December 2012, 1 727 individual samples, which were cryopreserved in Liaoning Cord Blood Bank on the basis of Rubinstein＇s method, were analyzed using blood counter to calculate TNC, colony forming unit （ CFC ） to find the hematopoietic cells, and flow cytometry to determine the CD34%. Another analysis was conducted to find the factors that influenced TNC and to elucidate the relationship among TNC, CFU-C and CD34 ~ CD45dim ±. The viability and functional capacity of the 23 cord blood sample which were cryopre- served over 24 months were evaluated. The thawed sample from segments, tubes and bags were assayed separately. Results The average volume of cord blood was 96.62 ± 20. 41 mL. The average TNC, total CD34 ~ and total CFC were （ 9. 88 ± 2. 54）× 105, （2. 76 ± 1.85） ×106 and （ 11.05 ±7.72） × 105, respectively. The relative ratios of TNC and total CD34 and total CFC were both 0. 49. The percentage of CD34 ~ and CFC/105 was 0. 80. After analyzing 23 cases of cord blood be- fore and after thawing, compared to fresh samples in bags, tubes and segment, the numbers of white cells （× 109/L） were 28.03 ± 8.01, 22. 55 ± 6. 56 and 20. 43 ± 5.23, respectively; the numbers of TNC （ x l0s ） were 10. 74 ± 3.33, 58 ± 2.90, 7. 78 ±2. 28, 7.97 ± 2.73; the tryphan blue stain resistance levels （%） were 99. 0 ± 0, 83.04 ± 6.21, 78.9 ± 6.56, 80.43 ± 6. 01 respectively. Compared with the fresh samples, the leukocytes count, TNC and tryphan blues stain-re- sistance decreased significantly （P 〈 0. 01 ）, so did the total numbers of white blood cells and the percentage of decrease of tryphan blue in segments and tubes （P 〈 0. 01 ）. As for the CFU-C production, the numbers

关 键 词：人脐带血 造血干细胞 细胞总数 CD34+ 集落形成单位(CFU-C) 活细胞冷冻保存 公共脐血库 质量控制 

分 类 号：R457.7[医药卫生—治疗学]