Does anyone know of a method/tip to avoid interstitial cell contamination from endothelial cells? I have been trying to isolate valvular endothelial cells (pig) and keep getting interstitial cell contamination.
Any help would be greatly appreciated!
Do you see this contamination right away or after a passage or two? Could you try MACs or FACs sorting using PECAM or VEcad either after your initial harvest or after your founding passage, and replate a purified population...... maybe trying different digest times to help elimante excess cells "tagging along" with the ECs"
https://www.ncbi.nlm.nih.gov/pubmed/19137801
Thanks for sharing this paper! Yes, I usually see the contamination few days after the isolation when the cells starts ti grow.
I haven't tried the other techniques using antibodies because I am planning to use these cells for long period of experiments. I am afraid that it will alter my results when I do gene expression.
You can try culturing on matrigel. Thus you can enrich the epithelial cells in your case the endothelium.
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