Here I am trying to optimizing the growth of breast cancer cell which is ++- requiring additional growth factors (20% FBS, insulin and EGF) for its growth. After adding the required growth factors in the media It doesn't grow and proliferated well. Have someone any kind of opinion What major factors are required for its growth?
You may also try higher than 5% carbon dioxide tension. It is known to work sometimes.
Thank you Sir. Other than that shall I use any different kind of growth factors?
Actually, there is a lot that can be tried. Many breast cancer cells exhibit estrogen-dependence to near-obligate extents. So, you may try adding varying concentrations of estrogen. You may also try some ECM coats on the substrate like fibronectin, etc. One important recommendation that I'd like to make is to protect your cells from aeration (oxygen) during handling.
I have used Matrigel as a substrate to grow primary cultures of breast cancer cells derived from patient's biopsies. At the start, without coating, some of them had a problem to grow, but with a coating, we were able to grow almst all of them. I have used Martrigel as it is more complex, than simply fibronectin, and because we had a lot of it leftover after other experiments. We had the kind with a low level of growth factors, so the main point was not the addition of growth factors, as there was enough of them in a medium, but a formation of a good niche for the growth of tumor cells, which were just moments ago sitting nicely in a beautifully formed niche by tumor fibroblasts.
If it was helpful for the primary breast cancer cultures, than it should also help with your difficult cell ine.
Even if you use just fibronectin, it should help, as it should be enough to stimulate integrin signalling and decrease anoikis together with stimulation of proliferation and anti-apoptotic pathways.
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