中国组织工程研究2012，Vol.16Issue(1)：56-60,5.DOI:10.3969/j.issn.1673-8225.2012.01.012
转白血病抑制因子基因人胚肺成纤维细胞对脐血CD34+细胞体外扩增的影响
Effects of human embryonic lung fibroblasts feeder layer transfected with leukemia inhibitory factor gene on the proliferation of umbilical cord blood CD34+ cells in vitro
摘要
Abstract
BACKGROUND: The number of hematopoietic cels in single copy cord blood is limited, and it is difficult to meet the needs of adults. How to effectively amplify cord blood hematopoietic stem/progenitor cels (HSPCs) is a hot res earch issue currently. OBJECTIVE: To construct gene-modified human embryonic lung fibroblast cels using human leukemia inhibitory factor (hLIF) and to observe the effect of trans genic cells on the amplification of HSPCs in vitro.METHODS: The feeder layer trancfected with recombinat adenouirus Ad-hLIF was established and objective gene was detected by using RT-PCR and ELISA. Umbilical cord blood CD34+ HSPC were separated using ma g n et io-a ctivated cellsorting and the purity was detected using flow cytometry. Homing ability of amplified HSPC in vitro was detected by transmembrane migrationRESULTS AND CONCLUSION: Trans genic feeder cels were constructed successfully, and the expression of objective gene was verified by RT-PCR and ELISA. After culturing with feeder layer cels for seven days, the number of CD 34* HSPCs in the group containing hLIF was 15.73 much more than that in the group without hLIF. The expression rate of surface adhesion molecules wasfeeder layer was significantly higher than that of the control group, with a better homing ability. The feeder layer cells of adenovirus vector transfected with hLIF gene amplify umbilical cord blood CD34+ HSPC effectively in vitro and delay differentiation; what more, the CD34+ cells retains a great homing ability after amplifying in vitro.关键词
人白血病抑制因子/饲养层/CD34+细胞/体外扩增/造血干/祖细胞分类
医药卫生引用本文复制引用
夏卫,郁心,姜健,田丽娜,陈瑶,缪竞诚..转白血病抑制因子基因人胚肺成纤维细胞对脐血CD34+细胞体外扩增的影响[J].中国组织工程研究,2012,16(1):56-60,5.基金项目
江苏省卫生厅科研基金资助项目(H200955) （H200955）
无锡市社会发展科技计划项目(CSZ00962). （CSZ00962）
