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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! smile.gif

-amyerspsu-

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.

-Rhombus-