Dear all,
we would like to transfect our clones of A549, HepG2 and BEAS-2B cells with a GADD45a luciferase reporter plasmid that will be sent from a research team in China.
This research team have recently developed the transfected cell lines (HepG2 an A549) (Xin et al., 2015. The development of GADD45 luciferase reporter assays in human cells for assessing the genotoxicity of environmental pollutants. Toxicol Mech Methods, Early Online: 1–7) but they are not able to send us the obatined cells due to the distance between China and Italy.
They offered us to send the reporter plasmid but at the moment we are not able to perform the steps that lead to the transfected cell lines we would like to obtain.
Someone can help us with the trasnfection? or do you know if these cell lines already transfected are available in commerce on in some research group?
We known that Gentronix have stable transfected lines with GADD45a but we need A549 and HepG2 cells as host!
Thank you
Dear Diego,
We obtained good transfection efficiency with both cell lines using PolyMag Neo and Magnetofectamine from OZ Biosciences (you can refer to Wang et al., BBRC, 2011 and MontRose et al, BBRC, 2014). To increase your transfection rate with these sensible cells, you can perform your experiments on dishes improving cell attachement such as cellbind (Corning) or advanced TC (Greiner) in reduced serum medium such as Opti-MEM.
I wish you good luck with your project
Flavie
A549 and HepG2 are indeed common cell lines, but they are different and require separate transfection setups (for A549: https://altogen.com/product/a549-transfection-reagent-lung-carcinoma-ccl-185/ and for HepG2: https://altogen.com/product/hepg2-transfection-reagent-hepatocellular-carcinoma/). I have not conducted the specific transfection that you have, but I would still suggest following pre-established protocols for transfection into the cell line you're using, with particular emphasis on the concentration of the plasmids you are transfecting.
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