Hi, I am starting to use HUVEC from cord for the first time. Despite I paid attention, after having centrifugated and put them in a t25 flask, after 24 hours there are clots and cells look suffering. Any suggestions? I used HUVEC at the second passage after isolation from humancord, and I simply transfered half of them from a t25 flask to a new one. Thanks in advance! Laura
Hello Laura Calvillo
I have attached a protocol for your reference. Have you followed the protocol similar to the one provided in the article below?
Article Isolation of Human Umbilical Vein Endothelial Cells (HUVEC)
Best.
Hi Laura, does your flask have any coating? Our lab uses fibronectin-coated flasks for HUVEC culture, maybe that can help.
Hi Valentina, thanks for your support. Yes, we use a very good coat. The problem seems to be the efficiency in separating cells with 5 ml pipettes after centrifugation and the incubation time in trypsin. I will do more tests in the week... Thanks again!
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