一株IL-11 cDNA转染的成纤维细胞体外促造血活性的鉴定

Hematopoietic promoting characters of a IL-11 cDNA transfected fibroblast cell

为了探讨IL－11基因修饰成纤维细胞的体外促造血活性及其在造血系统基因治疗中的应用前景．方法：将含IL－11cDNA的逆转录病毒载体转染小鼠成纤维细胞系NIH－3T3细胞，获得了高表达IL－11的亚克隆1E7，通过造血祖细胞集落形成实验及细胞增殖实验对1E7细胞在体外的促造血活性做了初步的鉴定．结果：1E7培养上清或1E7细胞作为饲养细胞，在体外培养体系中均能明显地促进骨髓细胞GM－CFU，Meg－CFU和Mix－CFU的形成及骨髓细胞增殖．结论：IL－11cDNA基因转染的成纤维细胞能持续、稳定地表达造血因子目的基因，在体外具有促造血祖细胞集落形成及增殖的活性，是造血因子基因治疗中较为理想的靶细胞．

In order to test the in vitro hematopoietic promoting effects and availability of IL-11 gene modified fibroblast cells in gene therapy. Methods: We transfected a retroviral vector pDoIL-11 containing IL11cDNA into mouse fibroblast cell line NIH-3T3 by LiPofectin transfection method. After selection and cloning,a subclone 1E7 which could secrete high level of IL-11 was obtained. The colony-forming and proliferative promoting effects of 1E7 supernatant and 1E7 cells as feed cells in hematopoietic progenitor cell culture were preliminarily identified by semi-solid colony forming experiment and liquid cell culture. Results: Both the culture supernatant of 1E7 and 1E7 cells as feed cells could significantly promote the colony formation of GM-CFU,MegCFU and Mix-CFU. Conclusion: IL-11 cDNA modified fibroblast cells can be used as feed cells in hematopoietic progenitor cell culture and can be an ideal target cells in the hematopoietic gene therapy.