I've been trying to perform wound healing assay on HUVECs, however, whenever they reach 90%-95% confluence they start detaching from the surface forming gaps in the culture making it impossible to have a 100% confluence covering the entire surface.
What is the best way to grow HUVEC to 100% confluence so they can be used for wound healing assay.
For cultivating the HUVEC cells I use this media and the cells like it a lot.
1% Pen.Strep/1% L-Glutamine/25mM HEPES/10% FBS / LSGS in Medium 200
LSGS
https://www.thermofisher.com/order/catalog/product/S003K
I recommend to you to use DMEM media by this component:glutamax (high glucose) [+] 1 g/l D-glucose, HEPES 15mM, pyruvate &NaHCO3+FBS 10%
In my experience HUVEC cells never reach 100% confluency, they start detachment after 90 confluency. As these cells grow in monolayer, as soon the cell to cell contact is established they start detachment. The best way is to perform your test at the maximum confluency you get.
It depends on what type of media you use. I use an endothelial cell media, you can either use EGM media from Lonza or ECM media from ScienCell. From my experiments, the EGM media allowed rapid growth but the ECM media allowed constant maintenance of the cells.
- Badges
- Science topic