Hu L, Zhang B
Department of Blood Transfusion, Xiehe Hospital, Tongji Medical University, Wuhan 430022.
J Tongji Med Univ. 2000;20(4):287-9. doi: 10.1007/BF02888181.
Liposomes have showed many advantages in mediating exogenous gene into many cell types in vitro and in vivo. But few data are available concerning gene transfer into hematopoietic cells. In this report, we described two-marker genes (Neo R and Lac Z) co-transferred into hematopoietic cells of human and mouse by using liposome in vitro. The efficiency of gene transfer was tested by X-gal staining and observation of colony formation. 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 G418 selection, the blue cell rate was (46.06 +/- 3.47)% in human and (43.45 +/- 4.1)% in mouse, which were markedly higher than those before selection, suggesting that high-efficiency gene transfer and expression could be attained in primary hematopoietic cells using this easy and harmless transduction protocol. At the same time, this protocol provided experimental data for clinicians to investigate the biology of marrow reconstitution and trace the origin of relapse after autologous bone marrow transplantation for the patients with leukemia.
脂质体在介导外源基因导入多种细胞类型的体外和体内实验中已显示出许多优势。但关于基因导入造血细胞的数据却很少。在本报告中，我们描述了利用脂质体在体外将两个标记基因（Neo R和Lac Z）共导入人和小鼠的造血细胞。通过X - gal染色和集落形成观察来检测基因转移效率。在未进行G418筛选的情况下，转导细胞的X - gal蓝色染色率在人约为(13.33±2.68)%，在小鼠约为(16.28±2.95)%。经过G418筛选后，人源的蓝色细胞率为(46.06±3.47)%，小鼠源的为(43.45±4.1)%，均显著高于筛选前，这表明使用这种简便且无害的转导方案可在原代造血细胞中实现高效的基因转移和表达。同时，该方案为临床医生研究骨髓重建生物学以及追踪白血病患者自体骨髓移植后复发的起源提供了实验数据。
