How can I obtain multicellular spheroids from MCF-7 human breast cancer cells.
You can seed 1000 cells per well in an ultra-low adherent plate in a serum free media and adding a cocktail of growth factors like EGF (10ng/ml), FGF (10ng/ml), Insulin (20ng/ml), and Lif1 (10ng/ml).
Be sure to change the media every 2 days. Spheroids will start forming in 4-5 days and can then be counted under a light microscope.
Hope this helps.
You can seed 1000 cells per well in an ultra-low adherent plate in a serum free media and adding a cocktail of growth factors like EGF (10ng/ml), FGF (10ng/ml), Insulin (20ng/ml), and Lif1 (10ng/ml).
Be sure to change the media every 2 days. Spheroids will start forming in 4-5 days and can then be counted under a light microscope.
Hope this helps.
If u do not have access to low adhesion plates u can use plates coated with ultra pure agarose which will avoid cell adhesion.
@ Apurva Atak
Thank you so much for the suggestion. I will try this method. Is does not need to add any kind of gelatin agent like CMC or other else ?
I advice you to use ultra-low attachment plates from Corning.They are more efficient than agarose plates.
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