人骨髓基质细胞促进脐血CD34＋细胞的体外扩增和对NOD／SCID小鼠的植入  

Human bone marrow stromal cells facilitate the cord blood CD34＋ cells ex vivo expansion and shortterm engraftment in NOD/SCID mice

作　　者：费小明[1] 吴雨洁[1] 常在[2] 缪扣荣[1] 周小玉[1] 潘芹芹[1] 汪承亚[1] 

机构地区：[1]南京医科大学第一附属医院,210029 [2]南京医科大学模式动物研究所

出　　处：《中华血液学杂志》2008年第2期97-100,共4页Chinese Journal of Hematology

摘　　要：目的研究人骨髓基质细胞（MSC）促进脐血CD34＋细胞体外扩增及植入能力的作用。方法分离、培养正常人的MSC作为滋养层细胞。在TPO、SCF、FL和G-CSF刺激下，比较有、无MSC滋养层细胞对扩增脐血CD34＋细胞后，CD34＋细胞和CFU数的增加倍数，以及植入非肥胖性糖尿病／重症联合免疫缺陷（NOD／SCID）小鼠的能力。结果以骨髓MSC为滋养层细胞的培养体系可以更加有效地扩增脐血CD34＋细胞。体外扩增l周总细胞数（TNC）、CD34＋细胞和CFU的均数分别增加111．6、19．3和58．0倍；体外扩增2周后TNC、CD34＋细胞和CFU的均数分别增加532．8、41．3和563．5倍。脐血细胞输入NOD／SCID小鼠6周后，移植未扩增脐血细胞的对照组小鼠骨髓细胞中人CD45＋细胞比例仅为1．2％～3．7％；移植单纯细胞因子刺激扩增组小鼠骨髓中，人CD45＋细胞比例为7．6％～12．1％；以MSC为滋养层扩增的脐血细胞移植组，人CD45＋细胞比例达到45．3％～59．1％。结论以人骨髓MSC为作为饲养层细胞，不但可以更加有效扩增脐血CD34＋细胞，而且可以促进脐血细胞植入NOD／SCID小鼠的能力，有潜在的临床应用价值。Objective To explore the potential of human bone marrow stromal cells （MSCs） as the feeding-layer to promote ex vivo expansion of cord blood CD34 ＋ cells and engraftment of the expanded cells in NOD/SCID mice. Methods Human MSCs were routinely isolated and cultured. MSCs at passage 3 were used as feeding-layer for the expansion of cord blood CD34＋ cell in the presence of thrombopoietin （ TPO ）, flt3/flk2 ligand（FL） , stem cell factor（SCF） and granulocyte-colony stimulating factor（G-CSF）. The engraftment potential between unexpanded and expanded cord blood cells transplanted into NOD/SCID mice was compared. Results The total nucleated cells （TNC） , CD34＋ cells and colony forming units （CFUs） in the MSC feeding culture were increased by 111.6-, 19.3- and 58-fold after 1 week expansion and 532.8-,41.3- and 563.5- fold increased after 2 weeks expansion respectively as compared with that in non MSC feeding culture. In transplant experiment, the percentage of human CD5＋ cells （45.3% -59.1% ） in bone marrow of recipient mice transplanted with the MSC feeding expanded cells was the highest in all the groups at six weeks after transplantation. Conclusion Human MSCs enhance CB CD34＋ cells in vitro expansion and their capacity of short-term engraftment in NOD/SCID mice.

关 键 词：胎血 造血干细胞 扩增 骨髓基质细胞 

分 类 号：R686[医药卫生—骨科学]