Hi I am trying to develop a protocol in which I can co-incubate cancer cells and platelets to study the effects of platelets receptors/activity on cancer cells. I wonder if I could get the same protocol to design my experiment with variation.
You can label platelets with GFP and cancer cells with RFP proteins and can seed them together in 1:1 ratio in common medium. Then u can study whatever function u want to study.
Another option is to culture epithelial cells in matrigel and add platelets on top (see http://www.nature.com/nmeth/journal/v4/n4/full/nmeth1015.html for a starting point), this would mimic the interaction between two cell types in tissue and also allow you some degree of separation of the two cell types at the end of your assay even without labeling.
There are also other, more targeted options depending on the type of assay you wish to perform (see for example http://ibidi.com/applications/cell-based-microscopy-assays/co-cultivation/)
I have done co-culture experiments in the past using Transwell permeable (3.0uM) inserts (available through corning - #3452). This way I can plate one cell line in the insert and the other in the well - it works quite well if you are hoping to study the effects of secreted factors from one cell line on the other. If you are doing this in a 6 well plate I would recommend 50,000 cells per chamber and then allowing some time for the cells to grow in co-culture - I would also set up controls to ensure that both cell lines are relatively stable in the media you intend to co-culture them in - changing medias can drastically alter the behavior of your cells. If your predicted results are dependent on direct cell contact, this is clearly not the appropriate choice.
Dear All,
Thank you very much for informative ideas. I will soon get intouch with your answers once I finish reading the suggested protocols.
Best Wishes!
- Badges
- Science method