Can 3D culture be used to differentiate a cell population potentially containing both non cancerous and cancerous cells? If there is a population of cells growing derived from primary tumour samples (clinical samples) could you expect only cancerous cells to establish in 3D culture? Can it be used as a method to confirm the cells (at least some of them) are cancerous within the population if the establish in 3D culture? (A way of confirming cancer cells are present without other analysis i.e. metaphase spreads?)
There are many types of "3D" culture conditions, but I have grown primary breast cells (non-cancerous) on top of matrigel beds and embedded within matrigel. So yes, non-cancerous cells can grow in that kind of 3D culture.
What you want is a soft agar assay. ONLY transformed/tumorigenic cells can grow without attachment to a substrate. Normal cells will eventually die in this assay.
Article The Soft Agar Colony Formation Assay
We tried to grow fibroblasts in 3D cultures, and the cells liked it:)
Also, it is possible to combine cancer and non-cancer cells together, in my case I tried fibroblasts + different types of cells, especially when cancer cells alone did not make 3D cultures (fibroblasts help a lot to make nice spheroids). Article Differences of statin activity in 2D and 3D pancreatic cance...
Organoids can be grown from healthy cells. so this is not a general way to distinguish between cancer and non-cancer. Depriving wnt signaling does the trick (for some cell types).
Non-cancerous cells can grow in 3D. We have grown matrix-free normal brain astrocytes and endothelial spheroids. However, the number of cells required to grow non-cancerous cells should be more than cancer cells. For e.g. you would expect spheroids made from 100k/mL cells cancer cells would be bigger than spheroids made from 300k/mL astrocytes.
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