造血干细胞的基因转移及其表达的实验研究

Experimental Studies on Gene Transfer into Hematopoietic Stem Cells and Its Expression

为探索一种安全、有效、简易的非病毒基因转移策略用于介导造血干细胞基因转移的可行性,本研究利用商品化的脂质体将两个标志基因(Neo R和Lac Z)共转导造血细胞。通过G418筛选、富集转导阳性细胞,观察了外源基因在小鼠原代骨髓细胞的基因转移率及其表达的稳定性。结果显示:通过脂质体介导,外源基因在小鼠骨髓细胞可获得有效的瞬时和稳定的表达。同时,经G418筛选,转导阳性细胞可得到明显的富集。提示利用脂质体这一非病毒载体个导造血干细胞基因转移的可行性。

In order to explore the feasibility of gene transfer into hematopoietic stem cells mediated by using a safe, high-efficiency and easy, non-viral transduction protocol, we have transduced two marker genes (Neo R and Lac Z) into hematopoietic cells mediated by lipofectin. After selection with G418, the transduced positive cells were riched. Then we observed the efficiency of gene transfer in primary bone marrow cells of mouse and the stability of gene expression. The results show that the high-efficiency of short-(transient) and long-term (stable) gene transfer can be obtained in the hematopoietic cells of mouse mediated by liposome. Moreover, the transduced positive cells can be riched by G418 selection. The results indicate the feasibility of gene transfer into hematopoietic stem cells mediated by liposome, a non-viral transduction protocol.