Lu L, Zeitlin P L, Guggino W B, Craig R W
Department of Physiology, Johns Hopkins School of Medicine, Baltimore, MD 21205.
Pflugers Arch. 1989 Nov;415(2):198-203. doi: 10.1007/BF00370592.
Lipofection, a recently-developed method for gene transfer, was tested in secretory epithelial cells. Lipofection facilitated both transient DNA transfection with plasmids containing the chloramphenicol acetyltransferase gene and stable transfection with a plasmid containing the neomycin resistance gene, which confers resistance to the antibiotic G418 (Geneticin). Gene transfer occurred efficiently in a rabbit kidney medullary thick ascending limb cell line and in primary cultures of rabbit tracheal epithelial cells. The method was also effective in Simian virus 40-transformed human airway cells isolated from a normal individual and from a patient with cystic fibrosis (CF). Cytotoxicity was minimal, particularly when the time of exposure to the lipofectin-DNA was limited to 3-5 h (less than 5% cell loss). Thus, the lipofection method is useful for gene transfer in a variety of secretory epithelial cells and should be ideal for studies of defective secretory epithelial cell function in CF.
脂质体转染是一种最近开发的基因转移方法，已在分泌上皮细胞中进行了测试。脂质体转染既促进了用含有氯霉素乙酰转移酶基因的质粒进行瞬时DNA转染，也促进了用含有新霉素抗性基因的质粒进行稳定转染，该基因赋予对抗生素G418（遗传霉素）的抗性。基因转移在兔肾髓质厚升支细胞系和兔气管上皮细胞原代培养物中有效发生。该方法在从正常个体和囊性纤维化（CF）患者分离的猿猴病毒40转化的人气道细胞中也有效。细胞毒性最小，特别是当暴露于脂质体-DNA的时间限制在3-5小时时（细胞损失少于5%）。因此，脂质体转染方法对于多种分泌上皮细胞中的基因转移是有用的，并且应该是研究CF中分泌上皮细胞功能缺陷的理想方法。
