Hello,
Thanks to
RG members who answered my previous question, I could learn that HUVECs
do not change to other Cells without special techniques which induce
change of cell type during cell cultures.
This time, I want to ask whether HUVECs can change into other types of cells
during steps prior to cell cultures.(Receiving umbilical cord from hospital and isolation) by my unrecognized mistakes or variables.
I conducted real-time pcr and detected vegfr1 and vegfr3 expressions
so I concluded that Cells I extracted from umbilical cord are endothelial cells.
However, I learned that Endothelial cells other than HUVECs also express
vegfr1 and vegfr3 so I want to know whether HUVECs, during steps mentioned above,(Receiving umbilical cord from hospital and isolation) can change into
other types of cells by my unrecognized mistakes or variables.
Thank you!
Hello Jinkyu,
for HUVEC isolation is very important to isolate only endothelial cells without fibroblast, mural cells ... . From my experience, if you follow a standard protocol you should get about 97-99% endothelial cells. They look like cobblestone cells with tight contact with each other (if confluence >50%). See link below.
https://www.google.com/search?q=huvec&biw=1920&bih=1075&tbm=isch&source=lnms&sa=X&ved=0ahUKEwiRvr7si_XLAhWF2R4KHYUjBCUQ_AUIBigB#imgrc=OV7PYknw8o9MQM%3A
For your experiments use HUVEC up to passage 6-8.
To comfirm that you have only endothelial cells make qPCR for CD31, VEGFR2 but it will not say how pure your endothelial cells. To quantify endothelial cells and other type cells you should use Flow Cytometry or Immunostaining for endothelial specific marker (e.g. CD31).
During isolation and growing HUVEC they are pretty staible (passage up to 6-8) and don't change into other type of cells.
Dear Evgenii Boriushkin,
Thank you for your great and informative answer!
I was impressed with your knowledge.
I would like to second Evgenii's answer. Staining with CD31 (PECAM) or CD144 (VE-cadherin) is usually a good indication that you are looking at ECs. Also, staining with Von Willebrand factor (VWF) for the endothelial specific storage organelles Weibel-Palade bodies can be a good control.
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