Dear colleagues,
Does anyone has an idea on the optimal time interval between splitting one endothelial cell culture passage and the next?
Thank you in advance.
Hi,
the time intervals between passages, irrespective on cell type, depend on the rate at which the cells proliferate and reach confluency. You should passage your cultures when the cells will cover about 80% of a dish surface. The lower density of cells you will plate then, the longer will be the time to the next passage. And vice versa. So you can create your own, optimized protocol for cell passaging. When we work with endothelial cells, we usually split the cultures using 1:4 ratio and sub-cultivate them once a week.
Good luck
It's highly dependent on the cell line that you are using. We work with several of them and they differ between each other. I. E, HUVECs have a 1:3 - 1:4 sub cultivation ratio and we generally passage them twice a week. MLEC are slower and we only passage them once a week.
Hi Mr Anchang. I grow my endothelial cells at a seeding concentration of 1x10^5 cells/ml for 2-3 days. But this highly dependent to your culture medium supplements as well. If you added endothelial growth factors, you can expect the cells to grow faster. Thus you will need to optimise your protocol - try seeding your cells at different seeding concentrations at the same type of plates/vessels, and observe their growth for few days. Then you can pick which seeding concentration that you need based on the time you have planned for your experiments. All the best.
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