I need to cultivate endothelial cells (ECs) under low or no glucose conditions.
Usually I culture ECs using endothelial basal media (EBM) or endothelial growth media 2 (EGM2). But I can't remove glucose from EBM or EGM2 media.
I am considering using DMEM-based media for this purpose, as there are various DMEM formulations available with different glucose concentrations.
However, I am unsure whether DMEM is suitable for EC cultivation.
If anyone has experience in cultivating ECs in DMEM, I would greatly appreciate it if you could share your insights and any specific considerations for preparing DMEM media for ECs.
Additionally, if there are any other or better ideas for creating culture media with low or no glucose concentration for ECs, apart from DMEM, please share that information. Possibly RPMI1640, MEM, or L-15?
Thank you in advance for your assistance.
Dear Dr. Jung Kyu Han
You may use DMEM:F12 (DMEM/F-12 is a 1:1 mixture of DMEM and Ham's F-12). Please refer to the links below for more information on the media.
https://www.elabscience.com/p-dmem_f12_glucose_free_-458199.html
https://biowest.net/l0091-dmem-f12-wo-l-glutamine-wo-hepes-wo-glucose/#:~:text=Developed%20initially%20to%20study%20hormonal,and%20L%2DGlutamine%2Dfree.
I have worked with HUVECs and used the following medium, the recipe of which is provided below.
DMEM/F12- (50:50)
2% heat inactivated FBS.
1 µg/ml hydrocortisone
5 ng/ml EGF
10 ng/ml FGF-2
20 µg/ml heparin sulphate
250 ng/ml insulin
100 U/ml penicillin
100 µg/ml streptomycin
For HUVECs, one may use low glucose as low as 5mM approximately since high glucose causes increased apoptosis. For HUVECs, I would not recommend using cells beyond passage 8.
Hope this works!
Best.
Thanks for your kind help and detailed explanation. This was so practical information. I greatly appreciate it!
If you are aiming to culture primary endothelial cells, you may use MCDB131 media supplemented with endothelial growth supplements.
Article Type of culture medium determines properties of cultivated r...
Dear Prof. Jung Kyu Han: The linked article would clarify your question.
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