I need to perform an experiment on Caco-2 trasfection. I know that these cells are not easy to be transfected, in particular it is difficult when the cells are differentiated. Has someone some experiencies with Caco-2 cells trasfections? If so do you have a protocol ?
we havent tried to transfect Caco-2 cells with LDL. however, just about every cell type expresses the LDL receptor or a member of the LDL-receptor related receptor proteins. you may want to try using purified LDL or even particially purified low density lipoproteins. and also, you can up-regulate the receptors with statins or by starving the cells in medium less FBS for 3-4 hrs. these methods have worked for us.
The only method that had worked so far was the CaPO4 method. Kills many cells but i've managed to make a stable cell line afterwards.
J Lipid Res. 2010 Jul;51(7):1704-18. Epub 2010 Feb 19
method is simple.
40 ug LDL protein - 5 ug DNA; incubate in PBS-MgCl2 for 30 mins;
we incubate HeLa's in DMEM, 3%FBS, and mevastatin for 24 hrs. gently rinse cells with PBS - 3x. add DMEM with LDL-DNA cocktail to cells. in HeLa's we observe uptake in mins and we've seen GFP expression as early as 30 mins after. we've tested several different cell types, not all same response. some we transfected over night to see GFP.
Magnetofectamine worked quite well for those cells see Guix S., J Virol., 2007; 81(22):12238-48.
Dear Alberto, you can please refer to the links below. These links can help you for better understanding of Caco-2 cell transfection.
1) https://tools.lifetechnologies.com/downloads/Human_colorectal_adenocarcinoma.pdf
2) http://www.atcc.org/~/media/Transfection%20protocols/TransfeX/Caco-2%20on%20letterhead%20final.ashx
3) http://www.bio-rad.com/webroot/web/pdf/lsr/literature/bulletin_5370.pdf
Hi Alberto, I hope you have succeded to transfect your Caco2 cells because huis question was asked 2 years ago!
Anyway, another successfully transfection of Caco2 cells with DreamFect Gold
http://www.ozbiosciences.com/transfection-dna/19-dreamfect-gold-transfection-reagent.html
http://pubs.rsc.org/en/content/articlelanding/2015/ra/c5ra06562c#!divAbstract
Good luck
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