人脐静脉内皮细胞支持体外造血和造血干/祖细胞扩增  

Human umbilical vein endothelial cells support hematopoiesis and expansion of hematopoietic stem/progenitor cells in vitro

作　　者：袁红丰[1] 郭子宽[1] 刘晓丹 张双喜[1] 吴英[1] 毛宁[1] 

机构地区：[1]军事医学科学院基础医学研究所,北京100850

出　　处：《军事医学科学院院刊》2001年第1期45-49,共5页Bulletin of the Academy of Military Medical Sciences

基　　金：国家"八六三"计划资助项目 !(BH 0 3 0 5 0 1)

摘　　要：目的 :研究人脐静脉内皮细胞 (humanumbilicalveinendothelialcell ,HUVEC)对体外造血和造血干 /祖细胞扩增的支持作用 ,为造血干 /祖细胞体外扩增提供新的实验资料。方法 :用甲基纤维素集落形成实验检测集落形成情况 ,采用HUVEC与脐血单个核细胞和CD34+细胞直接接触共培养的方法观察HUVEC对体外造血和造血干 /祖细胞扩增的支持作用。结果 :HUVEC上清具有集落刺激活性。将HUVEC与脐血单个核细胞直接接触共培养 ,结果表明HUVEC能维持脐血单个核细胞体外存活至少 4周。将脐血CD34+细胞与HUVEC直接接触共培养 ,动态地观察有核细胞扩增倍数、CD34+细胞扩增倍数、CD4 1a+细胞百分比和CFU GM扩增倍数的变化。结果发现 ,有核细胞扩增倍数、CD34+细胞扩增倍数和CD4 1a+细胞百分率均在第 3周达最高 ,分别为 (6 8.1± 14.8)倍、(6 .6± 1.4 )倍和 15.6 % ,而CFU GM扩增倍数则于第 2周达最高 ,为 (5.7± 2 .1)倍。结论 :HUVEC能支持体外造血和造血干Objective: To investigate the role of human umbilical vein endothelial cells (HUVEC) in supporting hematopoiesis and the expansion of hematopoietic stem/progenitor cells in vitro. Methods: According to the fact that HUVEC supernatant has colony stimulating activity shown by methylcellulose colony forming assay and HUVEC can maintain the survival of mononuclear cells for at least four weeks in vitro, CD34 + cells from umbilical cord blood were seeded with (HUVEC group) or without (control group) HUVEC monolayer. Every week cells were collected and counted, the frequency of CFU GM was measured by using methylcellulose colony forming assay, and the percentage of CD34 + and CD41a + cells was measured by flow cytometry. Results : In control group,all the CD34 + cells died in two weeks. However, in HUVEC group,most nucleated cells and CD34 + cells were expanded by 68.1±14.8 fold and 6.6±1.4 fold,respectively at the third week while CFU GM expansion reached its peak (5.7±2.1 fold) at the week 2. Moreover, the percentage of CD41a + cells was enhanced significantly, reaching a maximum (15.6%) at the week 3. Conclusions: HUVEC can support hematopoiesis in vitro and expand the hematopoietic progenitor cells and CD41a + cells in direct contact coculture. [

关 键 词：人脐静脉内皮细胞 CD34^+细胞 CD41a^+细胞 造血细胞 

分 类 号：R329.28[医药卫生—人体解剖和组织胚胎学]