I am using 4 chamber slides for seeding HEK-293T and HEK 293 and rat hypothema cell lines. I am using RPMI-1640 from SIGMA with 10% FBS, 1% Gutamax and P/S.
seeding cells also on 24 wells (1x10*5) the same as chamber slides.
The problem is cells gathered in the center of chmaber slides and after trasnfection, they are floating more than 60 % like a cloud of cells.
I also tried with DMEM-H but I found the same problem.
I would be please of any kind of suggestion or help to sole this problem
What reagents are you using for transfection? and were the cells already adherent before transfection? It is not totally uncommon for adherent cells to detach after transfection but to not reattach is a bit strange. 293 cells are pretty sticky cells too. What kind of surface is on the chamber slides? IBIDI sells several kinds from what I can see.
Hi John,
I use XTREME gene transfection reagent, and 0.5 ug plasmid DNA (3:1).
Chamber slide is a 4 well (wall removable) permanoux chamber slide from sigma.
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