脐血细胞生物学特性的研究

Study on biological characteristics of umbilical cord blood cells

目的 :研究单份脐血造血干 /祖细胞 (HSC/ HPC)的质和量 ,脐血中的干细胞、免疫细胞的表型特征。方法 :收集 2 0份脐血 ,采用 6 %的羟乙基淀粉沉淀去除红细胞 (RBC) ,利用甲基纤维素半固体培养法培养和观察 HSC/ HPC的集落生成情况 ,以了解增殖能力 ,用流式细胞仪检测脐血 CD34 + CD38- 、CD34 + 、CD34 + CD38+ 细胞量及免疫细胞的表型特性。结果 :2 0份脐血采集量在 4 0 m L～ 16 5 m L 之间 ,均数为 (78± 2 3) m L。每次收集的有核细胞 (NC)数在 4 .8×10 8～ 4 .6× 10 9之间 ,均数为 (1.4 3± 0 .96 )× 10 9,NC的回收率为 86 .6 %。脐血中 CD34 + CD35-细胞数占 NC总数的 (0 .10 2± 0 .0 70 ) % ,平均为 (0 .12 0± 0 .0 90 )× 10 7/份 ,CD34 + 细胞占 NC总数的 (1.2 9± 0 .31) % ,平均为 (1.4 2± 0 .92 )×10 7个 /份 ,CFU- GM为 (2 .5 4± 2 .0 2 )× 10 6个 ,BFU- E为 (1.4 1± 1.39)× 10 6个 ,CFU- GEMM(1.6 0± 2 .30 )× 10 5个 ,CD4 + T细胞表达 CD4 5RA为 (89.95± 7.86 ) % ,CD8+ T细胞表达 CD4 5RA为 (99.5 8± 3.4 6 ) % ,均显著高于成人外周血 T淋巴细胞 (P<0 .0 1) ,结论 :单份脐血的 HSC/ HPC含量可以满足体重较轻的 (<5 0 kg成人和儿童患者移植的需要 ,体重 ) ,5 0

Objective:The study is aimed to determine the quality and quantity of HSC and HPC (hematop oietic stem/progenitor cells) in per sample cord blood,immunophenotypic features.Methods:20 samples of umbilical cord blood were collected.RBC was separted by using 6 % HES.The capacity of colony forming unit of stem/progenitor cells was observed and analyzed by means of methylcellulose culture method.The immunophenotype of the HSC/HPS and lymphocytes was determined by FACS.Results:The amount of 20 samples of cord blood ranged from 40 mL to 165 mL,averaging (78±23)mL.The number of NC (Nucleated cell) collected every time was about 4.8×10 8～4.6×10 9,averaging (1.43±0.96)×10 9.The recovery of NC was 86.6 percent.The number of CD 34 +CD 38 - cells accouted for (0.102±0.090) percent of NC in cord blood,averaging (0.102±0.090)×10 7 per sample.The number of CD 34 +cells made up (1.29±0.31) percent of NC,averaging (1.42±0.92)×10 7 per sample.The colongy yield of CFU-GM and BFU-E and CFU-GEMM was (2.54±2.01)×10 6,(1.40±1.39)×10 6 and (1.60±2.28)×10 5 respectively.CD 34 +RA in CD 4 + T and CD 8 + T cells was determined as (89.95±7.86) percent and (99.58±3.46) percent respectively,Which is remarkably higher than the number of T cells in adult peripheral blood ( P <0.01).Conclusion:The HSC/HPC content in persample of cord blood is enough in transplantation of children and adults who weight less than 50 kilograms.HSC/HPC,however,must be expanded exviro before transplantation for adults above 50 kg Graft-versus-host disease occurs much less when cord blood cells are transplanted because of immature immunocytes and weaker immunity.Original HSC/HPC,enough proliferation.