We are planning to culture HUVECs. However, we have a large stock of poly-L-lysine coated flasks and everyone is suggesting to culture the cells on either fibronectin or gelatin coated flasks or well plates.
Before purchasing another stock of untreated flasks, we were wondering if it is possible to use the ones we already have for this purpose.
You can definitely give a try if you have enough HUVEC numbers.Gelatin is more widely used and the HUVEC propagate well on gelatin.I would suspect that PLL will slow down the HUVEC.
I would think that PLL and PDL are more known to support the ectodermal cell type(neurons,glia etc) but may not be efficient for the endothelial cells.
All the best!
We frequently make gelatin-coated coverslips for different assays in our lab, and we always coat with poly-l-lysine before adding the gelatin. Although that's not quite the same as having a pre treated flask, we've found that our coverslips actually work better with both.
Good luck!
Thank you all for your suggestions. we will definitely give it a try and let you know the result.
Robbert
I always use a Gelatin coat on poly-l-lysine coated 96-multiwell plate for my human aortic smooth muscle cells. And cells are ok! Good luck!
Poly L-lysine and Gelatin do not chemically react however, . with gelatin coating, you'll have surface variation, anyway, you'll only notice when you are scanning under the microscope. So, use a leveled surface when coating otherwise with surface difference you might get two layers of cells in an uneven area!
Thank you all for taking your time to answer the question. netxt week we will probably start an adapted procedure and I would be happyt o post the experience with it.
Robbert
hI Robbert, could you please share your feedback with us? I am facing the sam question right now... Cheers
Ana
Hi all, a bit late probably, but in the end after a lot of trial and error we found that the HUVEC are doing well on just the Poly-L-lysine coated flasks. No need for gelatine. Just the right culture medium (Basic vascular medium supplemented with the VEGF growth kit from ATCC).
By the way, thank you all for the suggestions. They gave us a direction to aim at.
Robbert
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