Lipofection of insulin-producing RINm5F cells: methodological improvements
2007 (English)In: Journal of liposome research, ISSN 0898-2104, E-ISSN 1532-2394, Vol. 17, no 2, 49-62 p.Article in journal (Refereed) Published
Cationic lipid/DNA-complexes have been widely used as gene transfer vectors because they are less toxic and immunogenic than viral vectors. The aim of the present study was to improve and characterize lipofection of an insulin-producing cell line. We compared the transfection efficiency of seven commercially available lipid formulations (Lipotaxi, SuperFect, Fugene, TransFast, Dosper, GenePORTER and LipofectAMINE) by flow cytometry analysis of GFP-expression. In addition, we have determined the influences of centrifugation, serum and a nuclear localization signal peptide on the lipofection efficiency. We observed that two lipid formulations, GenePORTER and LipofectAMINE, were able to promote efficient gene transfer in RINm5F cells. However, GenePORTER exhibited the important advantage of being able to transfect cells in the presence of serum and with less cytotoxicity than LipofectAMINE. LipofectAMINE-induced RINm5F cell death could partially be counteracted by TPA, forskolin or fumonisin β1. Finally, both centrifugation and a nuclear localization signal peptide increased transfection efficiency.
Place, publisher, year, edition, pages
2007. Vol. 17, no 2, 49-62 p.
RIN5mF cells, gene transfer, liposomes, cell death, NLS
Medical and Health Sciences
IdentifiersURN: urn:nbn:se:uu:diva-13101DOI: 10.1080/01676830701374986ISI: 000247781800001PubMedID: 17613696OAI: oai:DiVA.org:uu-13101DiVA: diva2:40871