Hello,
I am an graduate student who work with huvec.
Some days ago, I did wound healing assay with HUVECs.
After culturing the cells with 20% EGM, I starved HUVECs with approximately
1% EBM(50ml EBM + 500ul FBS).
I learned that antibiotics should be added to cell media.
I wonder whether I must add antibiotics to EBM mentioned above.
I did not add antibiotics to EBM.
(I also did not add antibiotics to EGM)
There was no contamination and I got an result as i expected.
But I want to make sure whether adding antibiotics to EBM is required.
Thank you!
In wound healing assays the presence or absence of antibiotics should not impact the final result. You should take into account that control cells and the other experimental points should be under the same culture conditions (both without antibiotics)
In my opinion standard antibiotics (like penicillin/streptomycin) are there only to prevent bacterial growth/contamination. Most likely they don't have much impact on the result in this assay. However, it's obviously best to be consistent with the protocol to get comparable results - either always use antibiotics or not. So, in my opinion antibiotics are not required.
Personally I have routinely used penicillin/streptomycin when culturing HUVECs and also used strictly sterile work procedures. Antibiotics won't prevent all microbial contamination and for example mycoplasma can go unnoticed unless specifically tested.
Thank you for all of your answers!
All answers are great help for me!
Thank you again!
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