脐血干细胞库的建立及其临床应用

目的建立无关供者脐血干细胞库,用于治疗骨髓衰竭、恶性及非恶性血液病、某些遗传病、重型免疫缺陷等疾病的干细胞移植.方法采集足月顺产或剖宫产的新生儿脐血,以羟乙基淀粉分离红细胞,DMSO/LMD冷冻保存.每份脐血常规检测细胞活率、有核细胞计数、祖细胞体外培养、CD34+细胞检测、HLA配型及病原学检测等.结果共采集2318份脐血,合格1240份,合格率为53.5%,合格脐血采集的平均体积(98.3±27.0)ml,分离后脐血有核细胞为(1.2±0.5)×109,回收率为(88.90±9.65)%.脐血细胞存活率为(97.00±2.61)%.CFU-GM数为(5.18±14.35)×105,BFU-E数为(10.35±11.55)×105,CFU-Mix数为(1.03±1.47)×105.脐血分离后CD34+细胞百分率为(0.5±0.32)%,细菌培养阳性4例,4例脐血CMV-IgM阳性.在广州脐血库的172例脐血移植登记患者中,HLA匹配4个位点以上的占91.12%.至2000年5月共提供9份应用于无关供者脐血移植,其中6例植入,植入成功率66.7%.结论脐血库的建立可使脐血能够较好地应用于造血干细胞移植治疗,建立脐血库有良好的应用前景.

Establishment of cord blood stem cell bank and its clinical application

OBJECTIVE: To establish unrelated cord blood stem cell bank for providing potential grafts for stem cell transplantation in the treatment of some malignant and nonmalignant disorders. METHODS: All cord blood units(CBU) were obtained from full term normal vaginal and Caesarean-section deliveries. Most of RBC was removed from CBU by the hydroxyethyl starch (HES) method. CBUs were cryopreserved in liquid nitrogen using DMSO and LMD as cryoprotectants. For each CBU, automated cell count, manual viability, HLA typing, screening for infectious agents, colony-forming assays and CD34 analysis by FACS were routinely performed. RESULTS: A total of 2,318 CBUs were collected, among which, 1,240 CBUs met all tests and standards. The average volume of collected qualified cord blood was (98.31 +/- 26.95) ml, the average amount of nucleated cells after separation was (1.22 +/- 0.47) x 10(9), and the average recovery percentage of nucleated cells (88.90 +/- 9.65)%. The average viability was (97 +/- 2.61)%, the number of CFU-GM (5.18 +/- 14.35) x 10(5), of BFU-E (10.35 +/- 11.55) x 10(5), and of CFU-GEMM (1.03 +/- 1.47) x 10(5). The average percentage of CD34+ cells after CBU separation was (0.5 +/- 0.32)%. Four units were positive in microbiological culture and 4 units CMV-IgM positive. In 172 patients registeredd for cord blood transplantation in Guangzhou Cord Blood Bank, the percentage of 4/6 HLA antigens matched units was 91.12%. Up to May 2000, 9 units had been used in unrelated cord blood stem cell transplantation and 6 of them were successfully engrafted. CONCLUSION: It suggested that cord blood bank could be useful in transplantation and the establishment of cord blood bank had a good prospect.