Cells in collagen gel - Growing mammalian cells in collagen gel (Oct/01/2004 )

Hi

I am growing MCF7 cells in collagen gel. I would like to grow the gel in small wells made of PDMS gasket on top of glass surface - 1mm deep and 5 mm diameter. I would like collagen to stick to glass robustly so that it would wash off.

I am using about 10 ul of collagen in each well and then add 10 ul drop of media on top of it. When I wash the media with PBS, the collagen detaches from the surface.

Does anyone know how I make collagen to adhere to the glass surface (or even polystyrene surface) stably?

Thanks,
Anand.

I would increase the collagen concentration. However, I've never tried to make collagen gels on glass surfaces. I regularly grow cells in collagen gels in 24-well and 6-well plates (plastic) and as long as the collagen is at high enough concentration, the collagen stays attached to the plastic. Also, you should consider increasing the amount of collagen gel mix you add to the well. The pH of the gel is also critical for good gel integrity, so consider adding more NaOH. Also, be gentle when pipetting and removing media (that's usually not an issue if the collagen has good integrity; but it could be critical for collagen attached to glass).