Hi everyone,
I am planning on working with some HUVECs (ATCC CRL-1730) in the future and I have not cultured these cells before. I would appreciate any tips for optimal growth of these cells (eg. should I be pre-coating flasks, how often to change medium etc.).
I am particularly curious to know how many passages people have found they can get to before the cell survival declines - I have heard that with primary HUVECs this number is quite low.
I also would like to transfect these cells. I have been working with HEK293Ts and doing Lipofectamine 3000 transfections but I don't think this will be so successful in the HUVECs. Has anyone done electroporation with these cells?
Thanks very much in advance!
Hello. I transfected my vector into HUVEC cells using PEI, and the result was relatively good.
Hi, huvec are primary cells, so care should be taken on counting the passages. They need the appropriate medium, that is available from several vendors. They attach well to plastics, so there is no need for coating. Below is a link gor appropriateanipulation. Good luck.
https://www.sigmaaldrich.com/IT/it/technical-documents/protocol/cell-culture-and-cell-culture-analysis/primary-cell-culture/human-umbilical-vein-endothelial-cells?srsltid=AfmBOopKYWdSIcbUsqZ6csKWW-gUiyjoOSaSxdlHLjt5BR8IPqGcHhIk
Passage number depends on your experiments. Don't let them get overly confluent as they will go into senescence and slow down. Be sure to split them at 80%, do not allow them to get to 90% or more.
If you want to do angiogenic analysis then you need to use them prior to passage 10 to form tubules. so freeze them out early and be ready to conduct your experiments when you thaw them.
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