Transfection of ECV304 Cells

[marc.bolliger at bc.biol.ethz.ch]

Hello

I need to transfect endothelial cells. For this reason, I chose the cell line
ECV304. Initial experiments showed that it is very difficult to transfect
endothelial cells. I tried the SuperFect and the Effectene transfection
reagents from QIAGEN but without big success (transfection efficiencies
around 0.05%). The systems worked well with COS-7 cells. The best results I
achieved with the CaPO4 method, but the efficiency is also unacceptable. Has
someone experience in transfection of ECs and would help me?

A second problem is that I hardly can make single cell suspension with the
ECV304 cells. They like to build aggregates. I tried trypsin/EDTA solution,
DNAse treatment, mechanical treatment through a melted pipette. Is it possible
that the number of passages is too high?

Thanks
Marc Bolliger
Laboratory for Biochemistry I
ETH Zurich
Switzerland

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