山东脐血库3000份脐带血处理保存的操作程序  

Operating Procedure of Collection, Processing and Preservation of 3000 Units Umbilical Cord Blood in Shangdong Cord Blood Bank

作　　者：周胜利 沈柏钧 阎文瑛 旭日 潘杰 马秀峰 宋道刚 

机构地区：[1]山东脐血造血干细胞中心,济南250002

出　　处：《中国实验血液学杂志》2001年第2期153-159,共7页Journal of Experimental Hematology

摘　　要：大量的实验和临床资料表明 ,脐带血干细胞移植可成功的用于造血干细胞重建。本文从30 0 0份脐带血的采集、分离、浓缩、低温保存、融化和各种检测等方面的经验 ,阐述如何在我国建立符合cGMP要求的高质量脐带血造血干细胞库。采用密闭式收集的脐带血和羟乙基淀粉 (HES)混合 ,浓缩有核白细胞 ,经过微生物及组织配型 (HLA)检测后 ,以液氮深低温保存供临床应用。贮存脐带血平均有核细胞 (NC)总数为 (1.2± 0 .6 )× 10 9,CD34+ 细胞为 (3.0± 3.7)× 10 6,CFU GM为 (1.1± 0 .7)× 10 6,CFU GEMM为 (1.1± 1.2 )× 10 6,回收率分别为 91% ,88% ,85 %和 82 %。红细胞和血红蛋白的回收率平均为 (39± 9) %和 (40± 8) % ,平均贮存体积为 (35 .1± 7.1)ml。从脐带血收集到分离的平均时间为 15小时 ,贮存前的脐带血活细胞比率大于 95 % ,冻融后的活细胞比率大于 92 %。单个核细胞 ,CD34阳性细胞和CFU GM的回收率分别为 96 % ,90 % ,和 91%。所收集脐带血中 ,未发现有人免疫缺陷病毒抗体 (HIVAb)阳性 ,梅毒阳性率为 0 .1% ,乙型肝炎表面抗原 (HBsAg)阳性率为 0 .8% ,乙型肝炎核心抗体 (HBcAb)阳性率为 3.2 % ,丙型肝炎抗体(HCVAb)阳性率为 0 .2 % ,母血巨细胞病毒 (CMV )免疫学检测阳性率为 87.1% 。The experience with the umbilical cord blood (UCB) stem cells for unrelated transplantation from our 3 000 UCB storage was described. UCB, collected from closed blood bags, were mixed with hydroxyethyl starch for nucleated cell (NC) enrichment. After finishing CD34 analysis, culture of hematopoietic progenitors (CFU GM and CFU GEMM) assays, microbial culture, HLA Class Ⅰ(A, B)serology and class Ⅱ(DR) low resolution SSP typing, cord blood units are stored in the liquid nitrogen for clinical applicatoin. Cord blood contained an average of nuclear cell(NC) (1.2±0.6)×10 9, CD34 + cells (3.0±3.7)×10 6, CFU GM (1.1±0.7)×10 6 and CFU GEMM (1.1±1.2)×10 6 for storage and the recovery rates were 91%, 88%, 85% and 82%, respectively. The recovery rates for red blood cell and Hb were (39±9)% and (40±8)%, respectively. The storage volume was (35.1±7.1) ml in a 50 ml storage bags. The mean time from collection to processing of 15 hours (range 4-24 hours) had no influence on cell viability. The cell viability before processing is more than 95% and 92% after UCB thawing. The recovery rates of NC, CD34 + cells and CFU GM post thawing were 96%, 90% and 91%, respectively. There were no HIV antibody (HIVAb) positive in all of UCB units. For an incidence of processed samples, infection with syphilis, HBsAg, HBcAb, HCVAb, CMV, bacterial contamination and abnormal hemoglobin were 0.1%, 0.8%, 3.2%, 0.2%, 87.1%, 1.2% and 0.1%, respectively. More than 3 HLA loci matched can be found for random patients in our cord blood bank and 6 HLA loci matched have 5%. For transplantation with nucleated cell counts of >2.7×10 7 cells/kg, our cord blood bank will be able to provide all of the umbilical cord blood stem cell samples for children and 50% of units can be used for some of adult recipients transplantation in the country. It is concluded that: ① The large cord blood banking for 20 000 UCB storage is feasible in China. ② Our system of whole procedure and methods is functionable for supplying qualified cord b

关 键 词：脐带血 脐血库 CD34^+细胞 造血干细胞 低温保存 造血祖细胞 

分 类 号：R457.12[医药卫生—治疗学] R331.1[医药卫生—临床医学]