Hello everybody,
Hope you all are doing fine during this difficult time.
I am trying to culture mouse oviductal epithelial cells but, the problem is when I culture them there is a mixture of epithelial and stromal cells. Does anyone have any idea how can I culture or separate epithelial cells only? I will appreciate any kind of input.
Thank you in advance!!
Stay safe!
Kind regards,
Seema
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Isolation and purification of epithelial and endothelial cells from mouse ...
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Please you look this publications
Article Establishment and Characterisation of Female Reproductive Tr...
This is what I do to separate epithelial cells from fibroblasts in my primary cell culture. I first try to mechanically remove the fibroblast if they have separate colonies from epithelial cells. If not, I trypsinize them first with 0.01% trypsin for 5 min at 37 degrees. Since epithelial cells are more sticky, they remain attached, and Fibroblast float. I then wash them with PBS and detach the rest of the cells with 0.25% trypsin/0.53 mM EDTA again 5 min at 37 degrees. After spinning and resuspending the cells in fresh media, I seed them in a culture dish and put them in the incubator for one to two hours. If there is any fibroblast left, they will attach first while the epithelial cells are still floating. I take the media and floating epithelial cells out and seed them in a new dish. I was always able to get a pure epithelial cell population using this method. I hope it helps.
Thank you so much Ms. Shahrzad Nouri for this valuable input. I will definitely try this protocol, I hope this works out for me too. Thank you so much again!!