Gene Transfer into Subcultured Endometrial Cells Using Lipofection
| Autor: | I. Lascombe, P. Mougin, C. Vuillermoz, G.L. Adessi, M. Jouvenot |
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| Jazyk: | angličtina |
| Rok vydání: | 1996 |
| Předmět: | |
| Zdroj: | BioTechniques, Vol 20, Iss 1, Pp 88-91 (1996) |
| Druh dokumentu: | article |
| ISSN: | 1940-9818 0736-6205 |
| DOI: | 10.2144/96201st03 |
| Popis: | Lipofection using the Lipofectin® reagent was optimized to transiently transfect subcultured guinea pig endometrial stromal cells with a β-galactosidase gene driven by a simian virus 40 promoter. Efficient transfection was obtained in the following conditions: a value of six for the ratio of lipofectin to DNA, a low cellular density (105 cells per 35-mm well) at the time of subculture (48 h before lipofection) and a lipofection duration of 12 hours. Lipofection was compared to calcium phosphate precipitation previously optimized in the same culture model. At a low cellular density, the lipofection method was found to be more efficient than the calcium phosphate precipitation. This result gives a great relevance to lipofection since the cultured cells available in an experiment are often limited. Then, using cells at low density and a plasmid containing the chloramphenicol acetyltransferase (cat) gene linked to an estrogen response element, it was shown that the lipofection procedure is a suitable tool for the evaluation of gene regulation by estrogen. |
| Databáze: | Directory of Open Access Journals |
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