转基因饲养层细胞体外促进脐带血CD34^+ HSPC增殖  

Transgenic cell line as a feeder layer on UCB CD34^+ HSPC in vitro amplification

作　　者：胡志清[1] 杨吉成[1] 盛伟华[1] 井莹莹[1] 苗莉[1] 张日[2] 朱南康[3] 缪竞诚[1] 

机构地区：[1]苏州大学医学部细胞与分子生物学教研室,江苏苏州215123 [2]苏州大学附属第一医院血液科,江苏苏州215006 [3]苏州大学放射医学研究所,江苏苏州215007

出　　处：《基础医学与临床》2009年第9期911-915,共5页Basic and Clinical Medicine

基　　金：国防科工委国防基础科研计划(A3820060130)

摘　　要：目的构建转hOSM基因细胞系作为饲养层细胞,观察其对脐带血CD34+HSPC的扩增作用。方法用分子生物学技术建立转hOSM基因细胞系作为饲养层细胞,免疫磁珠法分离脐带血CD34+HSPC,将其与饲养层细胞共培养7 d后,流式细胞术检测其增殖,将扩增后染色的CD34+细胞移植入SC ID小鼠体内,应用荧光显微镜和RT-PCR法观察移植效果。结果CD34+HSPC与饲养层细胞共培养后扩增了9.4倍,其表面归巢相关分子CXCR4和CD54阳性率仍较高,植入小鼠体内4周后,可成功归巢到小鼠骨髓。结论建立的转hOSM基因饲养层细胞可以有效地扩增CD34+HSPC,并维持其较高的移植效率和造血活性。Objective To establish the transgenic cell line of hOSM gene and observe its effect on the proliferation of umbilical cord blood （UCB） CD34^＋ HSPC as a feeder layer. Methods A transgenic cell line was established by molecular biology method and CD34^＋ HSPCs were separated by magnetic-activated cell sorting. After 7 day co-culture, the amplification effect was evaluated by flow cytometry. Then CD34^＋ ceils stained by CFDA SE were injected into SCID mice with sublethally irradiation and the effect of transplation was observed by fluorescence microscope and RT-PCR. Results The number of CD34^＋ ceils increased by 9.6 times and the expression rate of homing-ralated molecules was still higher than 50%. Four weeks after transplantion, CD34^＋ cells were found in bone marrow of SCID mice. Conclusion The transgenic cell line of hOSM gene as a feeder layer can effectively amplify UCB CD34^＋ HSPC ex vivo and can suppress its differentiation. Meanwhile, they have high transplantion efficacy and haematogenesis activity.

关 键 词：人抑瘤素M 反转录病毒载体 造血干/祖细胞 重度联合免疫缺陷病小鼠 

分 类 号：R457.7[医药卫生—治疗学]