Is it important to coat the flask / tissue culture plates etc with ECM like gelatin or collagen while culturing HUVEC or doing experiments with HUVEC. My cells adhere much better when I culture them on non-coated flask. I have my cells from LONZA, and I am using the EGM-2 media from LONZA as well.
I have cultured HUVECs with and without any coatings. These cells are not so sensitive as some microvascular EC which do not grow but die if there is no coating. You can also witness a drastic change in EC morphology with or without coatings if you look your cells under the microscope.
As a summary, I would recommend to to use gelatin/fibronectin even if HUVECs would not die with out it unless it affects to your experimentation in some more severe way. Use coatings still at least for stock culturing.
HUVECs do not need coating to adhere and grow. However, you may want to use coated plates for your final experimental stage as the cells tend to detach easily under some experimental conditions (e.g. multiple washings as in DHE staining or immunofluorescence studies). Otherwise, they should grow perfectly well in a standard tissue culture grade T25 or T75 flask without any coating.
Hi, ive cultured HUVECs without coating before and they attached just fine. Now I have thawed a vial and about half the cells didn't attach after 36 hours, though only some were dead. I have thrown out the supernatant. A good number of cells, many, that were not attached, still stayed in the flask, floating above the attached ones. Weird thing...
Any clues to explain this other than maybe having thawed a "bad vial" ?
Hello Virginia, I have exactly the same case. Please let us know if you find out what happend and I will do the same.
Best
Ana
Dear researchers,
Endothelial cells are polar cells with apical and basolateral domains which has to do with their attachments to the substratum, usually a basal lamina consisting of various types of collagen, fibronectin, laminin etc. Thus it is essential to grow them on a coated surface because glass surface is not known to these cells in their natural environment. I believe this will help us to deal with cells that are sane in our experiments.
Dear Virginia and Ana,
I noticed the same weird thing as well. Small amount of cells adhered, but the majority remained floating. And even after passaging, many detached and floated in the culture. Flow cytometry showed that at least 3% of the floating cells were dead, but they still remained CD31+. It´s weird and I still haven´t figured out the cause of this phenomena. Have you? I used the attached cells for further experiment.
Thanks for any suggestions.
Zuzana
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