获取稳定表达重组人血小板生成素(thrombopoietin, TPO)的NIH3T3成纤维细胞克隆,为开展成纤维细胞介导的 TPO基因治疗研究提供细胞学基础。方法:以重组人TPO cDNA为目的基因,构建真核细胞表达重组体pcDNA3/TPO,藉脂质体将其转染于NIH3T3细胞,经G418筛选,获得稳定表达重组人TPO的细胞克隆,应用PCR、RT-PCR及 Western印迹等技术鉴定目的基因的导入及表达。结果:pcDNA3/TPO可藉脂质体转染NIH3T3细胞,转染细胞可稳定表达并分泌TPO。结论:已获得所需细胞克隆,为应用人TPOcDNA进行基因治疗奠定了基础。 To obtain NIH3T3 fibroblast clone stably expressing recombinant thrombopoietin (TPO), and to provide a cytological basis for the study of fibroblast-mediated TPO gene therapy. METHODS: Recombinant human TPO cDNA was used as the target gene to construct recombinant eukaryotic expression vector pcDNA3 / TPO. The recombinant plasmid was transfected into NIH3T3 cells by lipofectamine. The recombinant plasmid was transfected into NIH3T3 cells by G418. , RT-PCR and Western blotting technology to identify the target gene of the introduction and expression. Results: pcDNA3 / TPO could be transfected into NIH3T3 cells by lipofectamine. The transfected cells could stably express and secrete TPO. Conclusion: The desired cell clone has been obtained, which laid the foundation for gene therapy using human TPO cDNA.