摘要
在介导体内外基因转移方面,脂质体已显示出许多优于病毒载体的特征.然而,有关脂质体介导造血细胞基因转移的报道很少.本文描述了脂质体介导双标志基因(Neo^R和Lac Z)在体外共转移进入人及小鼠原代造血细胞的研究.基因转移的效率通过X-gal染色及观察集落的形成来评价.结果显示,在存在G418的体系中,转导细胞能够存活并形成集落,而未转导细胞则不能存活.同时,转导阳性细胞能够被X-gal染成蓝色.其蓝染阳性细胞率在人及小鼠造血细胞于G418选择前分别为13.33± 2.68%和16.28± 2.95%.经G418选择启分别达到46.06±3.47%及43.45±4.1%,显著高于筛选前(P<0.01).提示,利用脂质体这一简单、安全的转导策略在原代造血细胞可获得高效的基因转移及稳定的表达.同时,共转导双标志基因进入造血细胞可用作高集转导阳性细胞.这一结论对于临床进一步探讨造血重建的生物学特征、追踪白血病的复发机制以及利用造血细胞作为靶细胞进行基因治疗的研究提供了实验依据.
Liposomes have several advantages over viral vectors for gene delivery, both in vitro and in vivo. However, few data arc available concerning gene transfer into hematopoietic cells. In this paper, we described the liposome-mediated two matker genes (Neo R and Lac Z) cotransfer into primary hematopoietic cells of human and mouse in vitro. The efficiency of gene transfer was measured by X-gal staining and observating the colony formation. The results showed that the transduced cells could be survival and develop colonies in the present ofG418, but the nontransduced cells could not live. In addition, the transduced cells could be stained by X-gal into blue. The X-gal blue staining rate of transduced cells was about 13.33± 2.68% in human and about 16.28±2.95% in mouse without G418 selection. After selection with G418, the rate of blue cells reached 46.06±3.47% in human and 43.45±4.1% in morse, which were markedly higher than before selection respectively. It suggested that high-efficiency gene transfer and experssion be attained in primary hematopoietic cells using this easy and harmless transduetion protocal and that cotransfer two marker genes into hematopoietic cells could be used to rich the transduced positive cells. It could provide a experimental basis for clinical workers to investigate the biology of mattow reconstitution, the relapse mechanism of leukemia and for gene therapy research taking bone marrow cells as targets.
出处
《现代临床医学生物工程学杂志》
1997年第4期235-237,共3页
Journal of Modern Clinical Medical Bioengineering
关键词
脂质体
标志基因
基因转移
造血细胞
Liposome Genetic marker Gene transfer Hematopoietic cell
