growing cells on coverslip - (Jul/17/2007 )
can i use normal coverslips to grow epithelial cells in 35mm(6well) TC plates?
i plan to coat them with collagen and fibronectin. I've seen some protocols on here about preparation but am wondering which is relevant/necessary. i'm only growing them for a few days before fixing them.
any help would be good as i have close to no experience in tissue culture.
when I have done this in the past, I coated with fibronectin and then overlaid with cell suspension, grew 2-3d, then fixed...it worked very well. I think the choice between fibronectin and collagen depends perhaps of your cell line? but I think that for most cases either will work
Depending on the cell line you may not even need to coat them.
The glass slips have a charge which allows many cells to cling to
them without a coat.
There are some coverslips that are treated with acid which makes the cells attach better. But for coating, just add the coating to the well and incubate overnight in 37 C. Don´t forget to wash in PBS before adding the cells (suspended in media), the cells should attach to the coverslips within 12 hours.
our cells are not very adherent so we use poly-D lysine and matrigel (for neurite outgrowth) before plating cells. One could use other extracellular matrix. Whatever suits the cells, is fine.
our cells stick to just about anything so we don't coat our coverslips but make sure you remember to autoclave the coverslips, or sterilize them somehow, before you use them.......I've made that mistake before!
we growed cells on normal coverslips, but we got better results with poly-l-lysine treated ones. Anyway, it is a bit of work to prepare them, so you should try first with non-treated coverslips.