Hello all,
I am try to grow LNCaP cells on BD Matrigel to perform some invasion and migration assays. However, my cells gets clumped as u can see in the attached in pic (they look fine on uncoated plates/wells) . Matrigel was diluted 1:100 and allowed to solidify for 3 hours in 370C incubator.
Any insights, tips or hints will be much appreciated.
Thank you
Shreyas
Even in cell culture medium in the plate or flask LNcap cells stick together. Most possibly due high level expression of adhesion molecules. Some conditions like acidic pH in the medium increases this characteristics. I think this help this type of cells to survive in a better way in the adverse condition.
You should change the cell density in Matrigel, and further, LNCaP cells should be spread in overlay medium containing 2% Matrigel and EGF. The direct embedded cancer cells in high density tend to exhibit the morphology shown in the picture.
Whilst I've never done a matrigel experiment with LNCaPs, in other experiments where I was trying to get as few clumps as possible I ended up passing the LNCaPs through a syringe (gently!) to make sure I wasn't plating clumps to start with, and to try and get them as single cells. Maybe that would help you here, at least you would be starting out with no clumps. This will mean they grow even slower though - they like to be able to touch other cells.
Dear All,
Thank you for all the inputs and apologise for delay in replying back. I will try seeding the cells in lower concentration as well as changing the matrigel concentration.
- Badges
- Science method