Fibroblats contamination in primary cell cultures - Fibroblats contamination in primary cell cultures (Sep/22/2006 )

Dear colleges:

We do lots of primary cultures and we have problems with fibroblats contamination. We had tried lots of think to make them disapear, but it was not possible.

So we are wondering how can we remove fibroblats from our cultures.

we do primary culture of epithelial cells, and preculture in an uncoated 75 ml flask where fibroblast adhere but not our epithelial cells; we take supernatant to culture epithelial cells, and have strongly diminished fibroblast contaminants

while culturing primary neuronal cultures, in order to avoid other cell types, we try to remove meninges as much as possible without much tissue damage.

Dear colleges:

We do lots of primary cultures and we have problems with fibroblats contamination. We had tried lots of think to make them disapear, but it was not possible.

So we are wondering how can we remove fibroblats from our cultures.

we grow primary epithelials and the medium itself is highly selective against fibroblasts. you might see as many as a dozen on a 10cm dish if you looked really, really close...not enough to hinder our experiments, for sure. however, once the cells are passaged too long or allowed to become too confluent, their growth slows dramatically and the few fibroblasts in the dish will actually begin to divide and grow if left alone for awhile

are you using early-passage cells in the correct medium, and passaging before confluence?

just use trypsin to get rid of fibroblast which is much more sensitive to trypsin than most epithelium. just repeat this for a few times and you will get pure epithelium in culture.

Fibroblasts attach to the substratum very fast and this characteristic of fibroblasts can be used to separate them from other cells. When you seed your primary culture in tissue culture flask or dish, after 20-25 min remove the medium from that flask and add it in to another one. If you see your first flask under microscope, you will find fibroblasts attached to it.

Hope it helps.