Hello All,
I just started using HepG2 cell lines and I would like to know the culturing conditions. I have read through the ATCC guidelines for this cell line and they recommend EMEM+10% FBS. However, I don't have EMEM and I read through literature where they have mentioned that even DMEM can be used. Unfortunately, the cells are growing very slowly in DMEM ( does anyone has tried using DMEM+10% FBS and faced similar issues ?)
How much time does HepG2 initial passage take to get confluent?
I would appreciate if someone can share few culture pictures of HepG2 ( without any treatment).
Can we also use DMEM F-12 for this cell line?
Best regards,
Ankur
Hi Ankur,
Yes DMEM is okay, I cultured HepG2s with 10%FBS in DMEM
I usually split them 1:5 or 1:6 and they will achieve confluency in ~3days. This will vary a little depending on how happy your cells are.
There is another thread on this with some photos:
https://www.researchgate.net/post/Can_someone_help_with_HepG2_cells_morphology
A,B,D are 'acceptable' morphologies for HepG2s
hope this helps,
ZQ
We would use DMEM and 10%FBS and found growth to be slow compared to other tumour cell lines, especially if they are seeded thinly as they like having other cells about.
We are very success growing Hep-G2.
Our HepG2 is growing at 37°C, 5%CO2 in a complete medium consist of the 500ml MEM alpha medium with GlutaMAX (life technologies 41090-028); 50ml FBS (no specific FBS required); 5ml Antibiotics/ -mycotics (100X) Gibco 15240-062; 5ml Sodium Pyruvate 100mM, (100X) Gibco 11360-039; 5ml MEM Non-essential Amino Acids Solution (100X) Gibco 11140-035.
For Washing solution, we use either D-PBS without Calcium/Magnesium.
For trypsinization , we remove medium from culture flask; wash cells pre-warmed D-PBS without Ca/Mg; prepare Trypsin solution 1X and deactivate the trypsin solution with D-PBS; centrifuge 1400 rpm at room temperature; resuspend the cell pellet in complete culture medium and culture in desired culture conditions; check cell culture and confluence on a regular base changing medium every 72hours.
For amplification, we split the cells ibn a ratio 1:2 or 1:3
For freezing, we prepare a freezing medium with a complete medium plus 4% DMSO and add 1ml Freezing Medium to the pellet, resuspend and transfer to cryo-vials and store them in a frosty freezing container at -80°C overnight and then store in liquid Nitrogen.
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