+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Lipid-mediated transfection of normal adult human hepatocytes in primary culture

, : Lipid-mediated transfection of normal adult human hepatocytes in primary culture. Analytical Biochemistry 247(1): 34-44

The aim of this work was to develop a procedure for the lipid-mediated transfection of DNA into normal adult human hepatocytes in culture. Cells were plated in a serum-free culture medium at various cell densities, on plastic or collagen-coated dishes, both in the absence and in the presence of epidermal growth factor (EGF). The cells were incubated for various periods of time with mixtures of DNA-lipofectin or DNA-3-beta(N-(B',N'-dimethylaminoethane)-carbamoyl) cholesterol (DC-chol) liposomes, and the efficiency of transfection was assessed by measuring the activity of reporter genes, beta-galactosidase or chloramphenicol acetyltransferase (CAT). For comparison, similar experiments were carried out with human cell lines including HepG2, Caco-2, and WRL68. The efficiency of transfection (in percentage of cells) was not significantly different after transfection with lipofectin or DC-chol and comprised between 0.04 and 1.7% (extreme values) for different cultures. The efficiency of transfection decreased as the age or density of the culture increased and increased in cultures treated with EGF. Direct measurement of the rate of DNA synthesis suggested that the efficiency of transfection was related to the number of cells entering the S phase. Under the same conditions, the efficiency of transfection was one to two orders of magnitude greater in the three cell lines. A plasmid harboring 660 bp of the 5'-flanking region of CYP1A1 (containing two xenobioytic enhancer elements) fused upstream of the promoter of thymidine kinase and the CAT reporter gene was constructed. When this plasmid was transfected in human hepatocytes, CAT activity was induced as expected. We conclude that normal adult human hepatocytes can be transfected with exogenous DNA and that the transfected construct is regulated in the manner expected from in vivo studies.


PMID: 9126368

DOI: 10.1006/abio.1997.2025

Other references

Higginson, J.; Oettle, A.G., 1961: Gastro-intestinal cancer in Africa, south of the Sahara. Acta - Unio Internationalis Contra Cancrum 17: 333-338

Rowe J.W.; Tobin J.D.; Rosa R.M.; Andres R., 1980: Effect of experimental potassium deficiency on glucose and insulin metabolism. Disturbances in glucose and insulin metabolism frequently accompany a variety of clinical states associated with K deficiency. The exact role of the K deficit and the mechanism of its effect are in doubt. The glucose-clamp technique was therefore...

Cousserans, J., 1980: Protected zones and mosquito abatement. The potentially conflicting interests of those conservationists concerned to establish and safeguard nature reserves and those with a responsibility for mosquito abatement, especially in areas of tourist attraction, are reviewed and briefly discus...

Jansen, T.L.Th.A., 2006: Atorvastatine for chronic synovitis due to massive intra-articular cholesterol monohydrate deposition in long-standing rheumatoid arthritis. In order to confirm the efficacy of ciprofloxacin (CPFX) incorporated into liposomes (CPFX-liposomes) for treatment of respiratory intracellular parasite infections, the influence of particle size on drug delivery to rat alveolar macrophages (AMs)...

Errington M.L.; Lynch M.A.; Bliss T.V.P., 1987: Long term potentiation in the dentate gyrus induction and increased glutamate release are blocked by d levo aminophosphonovalerate. D(-)Aminophosphonovalerate, a specific antagonist of the N-methyl-D-aspartate subtype of glutamate receptor, was perfused through a push-pull cannula into the dentate gyrus of rats anaesthetized with urethan in order to observe its effect on the i...

Abrahams, V.M.; Cambridge, J.; Lydyard, P.M.L.; Edwards, J.C.W., 1998: FcgammaRIIIa mediates tumour necrosis factor alpha secretion by adhered human monocytes. Immunology 95(SUPPL 1): 49, Dec

Sharma, U.; Chang, E.W.; Yun, S.H., 2008: Long-wavelength optical coherence tomography at 1.7 microm for enhanced imaging depth. Multiple scattering in a sample presents a significant limitation to achieve meaningful structural information at deeper penetration depths in optical coherence tomography (OCT). Previous studies suggest that the spectral region around 1.7 microm...

Bindelglas, P.M., 1973: "Conclusive evidence" and marihuana. British Journal of Addiction to Alcohol and Other Drugs 68(1): 51-56

Sokolov, S.S., 1956: Healing of wounds in radiation sickness. Vestnik Rentgenologii i Radiologii 31(1): 30-41

Schnitman, S., 2014: Endocrine glands and bones. Revista Odontologica 38(4): 114-133