逆转录病毒是最常用的基因治疗载体，但对人类细胞的基因导入率较低。本研究建立了一个流动转染体系，利用电动流速调节装置控制带有重组逆转录病毒的培养液，以一定速度缓慢流过生物半透膜，迫使病毒与在膜上生长的K562细胞充分接触，从而提高基因导入效率。经半固体培养NeoR基因/G418抗性集落法测定，无须筛选，单次基因导入率达80%，与常规静止转染法相比，差异十分显著（P＜0.01)。该装置简便、高效，可广泛地应用于基因治疗的基础和临床。%Retrovirus (RV) is the most widely used vector for gene transfer, but its efficiency is very low for human cells. A Flow-through Transduction System made in General Hospital of PLA was used to deliver interleukin-12 RV vector into human leukemia cell line K562. Compared with static procedure, the efficiency could be significantly improved,and detected by clony forming units (CFU) for NeoR gene in semisolid culture medium with G418 (83.1%±3.9% vs 9.2%±2.3%, P＜0.01). No significant difference was found in the levels of IL-12 p70 protein expression detected by ELISA, by flow-through transducted cells without G418 selection and by statically-transducted cells with G418 selection (11.3±7.5ng/ml vs 12.0±6.4ng/ml, P>0.05), while statically transducted cells without G418 selection could express very low IL-12 (0.1±0.024ng/ml，P＜0.01)。The Flow-through Transduction System is efficient and easy for use, it can be widely used in research and clinic of gene therapy.
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