脂质体介导骨髓细胞双标志基因转移与体外扩增研究

Liposome mediated gene transfer into bone marrow cells and its in vitro expansion

目的：探讨脂质体介导骨髓细胞基因转移的效率及其表达的稳定性，为基因治疗与基因标志研究提供实验依据。方法：利用脂质体介导，将ＮｅｏＲ和ＬａｃＺ两个标志基因共转导骨髓细胞，通过Ｇ４１８筛选、富集转导阳性细胞，然后扩增这些细胞，观察基因转移率及扩增对转导细胞基因表达稳定性的影响。结果：脂质体介导的骨髓细胞基因转移率为１３．３３％±２．６８％，经Ｇ４１８筛选、富集后，转导阳性细胞可达到４６．０６％±３．４７％。经体外扩增处理，扩增前后无显著性差异。结论：利用脂质体介导的基因转移方法介导骨髓细胞基因转移可获得高效、稳定的表达。

Objective:To explore the efficiency of liposome mediated gene transfer into bone marrow cells and the stability of its expression.Methods:Two marker genes (Neo R and Lac Z) mediated by liposome were co transferred into bone marrow cells.Following G418 screening,the positive cells were enriched and expanded in vitro.Results:The liposome mediated gene transfer rate in bone marrow cells was 13.33±2.68%,and after G418 screening, it was 46.06±3.47%.The blue colouring rates with X gal staining of these cells were not significantly different before and after expansion.Conclusion:Liposome mediated gene transfer in bone marrow cells is highly efficient and the expression of the transferred gene is stable.