I am currently working with primary human retinal microvascular endothelial cells (HRMECs) and need to grow tight confluent monolayers for TEER, transwell permeability assays, and TJ/AJ immunostaining. At the moment, my cells appear to grow in a 'patchy' manner, with some areas of high density and others where few, if any, cells grow at all.
Does anyone know of any ways which might result in growth of uniform, tight, confluent monolayers? If I leave the HRMECs in culture too long then they just start to die, rather than reach confluency.
I know some of these primary endothelial cell types have issues with contact inhibition and such. Is there any way to get around this?
Any advice would be greatly appreciated!
If you have the resources, you could try printing them: https://doi.org/10.1002/adfm.201801331
Otherwise here's a transwell assay reference: https://doi.org/10.1186/s12987-016-0035-0
Hello, when we culture primary porcine or rodent blood brain barrier cells, we
use from day 3 on culture medium containing 550 nM hydrocortisone, which results in higher TEEr values and morphologically better monolayers.
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