胚胎成纤维细胞与含LIF的培养体系支持鼠造血祖细胞体外扩增  

Expansion of Marrow Hematopoietic Progenitor Cells ex vivo Supported by Culture System from Mouse Marrow Embryonic Fibroblasts and Leukemia Inhibitory Factor

作　　者：袁国林[1] 邹萍[1] 程范军[1] 刘凌波[1] 伍晓菲[1] 王红祥[1] 

机构地区：[1]华中科技大学同济医学院附属协和医院血液病研究所,武汉430022

出　　处：《中国实验血液学杂志》2004年第6期807-811,共5页Journal of Experimental Hematology

基　　金：海外青年合作研究基金资助编号 :3992 80 10

摘　　要：为了探讨胚胎成纤维细胞与含白血病抑制因子 (LIF)的培养体系对鼠骨髓造血祖细胞体外扩增效应 ,并观察其对造血表面归巢相关黏附分子VLA 4 (CD4 9d)、VLA 5 (CD4 9e)、LFA 1(CD11a)、HCAM (CD4 4 )、L selectin(CD62L)的表达影响 ,将丝裂霉素C灭活的鼠胚胎成纤维细胞 (MEF)和含LIF的培养体系与鼠骨髓单个核细胞(BMMNC)体外共同培养 7天 ,检测BMMNC细胞总数、CFC、Sca 1+细胞百分率、细胞凋亡率和Sca 1+细胞表面黏附分子CD4 9d、CD4 9e、CD11a、CD4 4、CD62L的表达率。结果表明 :MEF和含LIF的培养体系明显扩增了BMMNC细胞总数、CFC和Sca 1+细胞 ,抑制细胞凋亡 (P <0 .0 5 ) ,而去掉MEF和LIF后的对照组培养体系BMMNC细胞总数明显下降 ,CFC和Sca 1+细胞完全死亡 ,细胞大部分凋亡 (P <0 .0 1)。扩增后Sca 1+细胞表面各黏附分子CD4 9d、CD4 4和CD61L表达与扩增前相当 (P <0 .0 5 ) ,而CD4 9e和CD11a表达明显高于扩增前 (P <0 .0 5 )。结论 :胚胎成纤维细胞和含LIF的培养体系体不仅可以体外显著扩增鼠骨髓造血祖细胞 ,而且扩增后的造血祖细胞(HPC)总体上保持其表面归巢相关黏附分子的表达 ,扩增后HPC归巢功能不受影响。The objective of this study was to explore the effect of culture system from embryonic fibroblasts and leukemia inhibitory factor (LIF) on expansion of mouse bone marrow hematopoietic progenitor cells ex vivo , and to observe its effect on the expression of homing related cell adhesion molecules among VLA 4(CD49e), VLA 5(CD49e), LFA 1(CD11a), HCAM(CD44) and L selectin (CD62L). The culture system from the mouse embryonic fibroblasts inactivatd by mitomycin C and contained LIF was used to culture with mouse BMMNC for 7 days. The total number of BMMNC, CFC, Sca 1 + cells, cell apoptosis rate and the expression of above cell adhesion molecules were counted. The results showed that culture system consisted of embryonic fibroblasts and LIF significantly enhanced the total number of BMMNC, CFC, Sca 1 + cells, suppressed cell apoptosis ( P <0.05). In control without MEF and LIF, the total number of BMMNC was reduced remarkedly, CFC and Sca 1 + cells were completely dead, the majority of cells produced apoptosis ( P <0.01). The expression of CD49d, Cd44 and CD61L on Sca 1 + cells were similar to that befor expression ( P <0.05), but the expression of CD49e and CD11a on Sca 1 + cells were remarkably increased ( P <0.05). It is concluded that culture system from embryonic fibroblasts and LIF can only significantly expand mouse bone marrow hematopoietic progenitor cells ex vivo , but the expanded hematopoietic progenitor may well sustain the expression of homing related adhesion molecules. The homing functions of these expanded hematopoietic progenitors kept no change.

关 键 词：体外扩增 造血细胞 胚胎成纤维细胞 白血病抑制因子 细胞黏附分子 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]