Abstract:
:We determined optimum conditions for delivering DNA to transformed human bronchial epithelial cells expressing wild-type (BEAS) or abnormal (2CF) cystic fibrosis transmembrane conductance regulator (CFTR) using cationic liposomes (Lipofectin, [N-(N,N-dimethylaminoethane)carbamyl] cholesterol[DC-Chol]/dioleoylphosphatidylethanolamine[DOPE], or LipofectAMINE) and reporter genes which measured overall transgene expression (luciferase) or the fraction of cells transfected (heat-stable alkaline phosphatase). All liposomes showed dose-related toxicity. Optimal liposome and lipid: DNA ratios were different for BEAS than for 2CF cells. For all 3 liposome preparations, small particle size and net cationic charge related to transfection efficiency. Both LipofectAMINE and DC-Chol/DOPE transfected a maximum of 3% of BEAS cells, but luciferase expression could be increased without increasing the fraction of cells transfected. LipofectAMINE transfected a maximum of 6% of 2CF cells, and luciferase expression could be increased with no further increase in fraction of transfected cells. DC-Chol/DOPE transfected over 12% of 2CF cells with relatively small increases in luciferase expression. We conclude that an optimal cationic liposome and lipid: DNA ratio for transfecting bronchial epithelial cells depends on: (1) small particle size and net cationic charge, (2) whether the cells have the cystic fibrosis defect, and (3) whether the desired outcome is transfection of the maximum fraction of the cells or maximum total expression of the transgene.
journal_name
Exp Lung Resjournal_title
Experimental lung researchauthors
Peters MT,Brigham KL,King GA,Meyrick BO,Gao X,Stecenko AAdoi
10.1080/019021499270259subject
Has Abstractpub_date
1999-04-01 00:00:00pages
183-97issue
3eissn
0190-2148issn
1521-0499journal_volume
25pub_type
杂志文章abstract::Investigations on leukocyte populations in the lung have shown that lymphocytes are found in different anatomical compartments. Lymphocytes can be seen to a different extent in the lung interstitium, the epithelium and lamina propria of the bronchi, the bronchoalveolar space, and the marginal lung vascular bed. Previo...
journal_title:Experimental lung research
pub_type: 杂志文章
doi:10.3109/01902149609070037
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journal_title:Experimental lung research
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journal_title:Experimental lung research
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更新日期:2020-01-01 00:00:00
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journal_title:Experimental lung research
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更新日期:1990-05-01 00:00:00
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