growing after filling in monolayer, please help - How and which genes/growth factors do it (Dec/10/2007 )
Hi all,
I am very curious about one problem. I am working now with known breast cancer cell line MCF-7. And with estrogen in medium ( non charcoal-treated serum) cells continue to grow after filling the bottom of the wells. I know it is very general question and I might as well google it, but I am quite an outsider in growth factor field, so it wasn`t so easy for me.
And here I would please you to help me with some tips on which proteins might be involved in this conus, or 3D- growth. I suppose there are people here working in this area.
Thanks for all answers in advance.
Andy
Did you mean that you want the cells further grow into 3-D instead of monolayer? Not every cell line does that due to contact inhibition. For 3-D, there are several matrix you can make yourself or you can buy from commercial sources.
Hi genehunter,
thanks for your responce!
I am sorry if I explained it wrongly. I meant that MCF-7 cell line indeed growth further into 3d, there is not stop or contact inhibition. And I was wondering what is behind it, what factors help cells to overcome contact inhibition, and, especially, what proteins are involved in that overgrowth.
i also culture MCF-7 but i don't use any estrogen, i use simple cdmem
i have never faced such a problem
i have never faced such a problem
Actually it is not the problem at all. And cells grow quite well when u use normal, I mean medium with all growth factors. But as MCF-7 is a cell line of estrogen dependent breast cancer, so after depleting estrogens from medium and using plasticware with no estrogenic compounds leakage, you will actually have cells stopping growing. And as it is difficult to achieve, cells will still grow, to fill you wells. And no more, they will not grow any further, due to contact inhibition as was told above.
But in normal medium or with defined estrogens, after 10 days you can see great conuses, or some structurees from the cells growing on each other. And I was asking about that case. What force cells to grow even though growth inhibition through contacts of the cells must occur?
A
As you probably know, MCF-7 cells are cancerous. That means they possess quite a lot of mutations/amplifications/deletions that dysregulate their growht. The cell machinery is always in full throttle, so the growth in 3D should be expected.
And as for the contact inhibition, the signaling cascade involve in this process is probably impaired, so even if the cells touch, they won't stop growing.