I am culturing liver cancer cell line in serum free tumour sphere medium (DMEM/F12 + EGF + bFGF) and culturing them for at least 7 days to enrich for cancer stem cells. Do I need to change the medium everyday?
I have a concern that this will disturb the sphere formation as I will need to centrifuge and I am afraid that during the resuspension in fresh media will affect the spheres. I am culturing them in 6-well ultra low plate by the way.
Hello,
You really don't need to change the medium every day, but it depends on the amount of spheres that are current in the well. We've cultured cancer stem cell spheroids in Petri dishes, the centrifugation did not affect their stability or behaviour. And the addition of EGF and bFGF is unnecessary.
We culture neurospheres from human glioma stem cells at cell density of about 100 000 cells/ml in neurosphere medium containing BIT, EGF and bFGF for 7 days without medium change. Passaging is made after low spin centrifugation followed by accutase digestion of the pellet. We use standard T25 or T75 flasks, as in the absence of serum these cells do not attach to the substratum. If medium change is required (acidic pH), spheroids are spun down at low speed and delicately transfered into fresh complete medium.
Hi Ain, you definitively don't need to change medium every day! A good approach to maintain your Tumorspheres in good helath and easily change medium is gravity sedimentation of the tumorspheres. Allow the spheres to settle by gravity sedimentation for 10 minutes at room temperature. Aspirate the supernatant, but leave approximately 200 μl in the conical tube. For more information in media, background and protocols - Have a look here: http://www.promocell.com/app/uploads/product-information/manual/CSC_Medium_TFE.pdf
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