Techniques for limiting fibroblast growth - Limiting fibroblast growth in keratinocytes (Sep/06/2006 )
Im getting a lot of unattached rounded up cells in my keratinocyte cultures so Im going to try washing the cells after a certain period of time to wash away the unattached cells from the plate/cover slip. Does anyone have any idea what time intervals I should use or how long I should expect the cells to attach to the plates? Any suggestions would be greatly appriciated, thanks!
We isolate smooth muscle cells and try and limit the Fibroblast contamination by differential plating.
The fibroblast's stick first and we do 4 x 15 minutes attachments, wash and keep the floaters. The eventual cells that are attached are characterised by alpha actin immuno staining and are 99 % SMC. Further sterile cell sorting removes the 1% Fibroblast's that are left.
In your culture, you will have to optimise the attachments times and characterisation.