Hi, does anyone know ho wto prepare conditioned medium (CM) and add to other cells.
A. This is what I understand based on what I've read:
1. Grow cells in 10cm dish
2. after 3 days when cells have reached 70-80% confluence, collect medium
3. centrifuge collected medium at 1,000 g for 10mins to remove cell depris etc and store in 1ml aliquots in -80deg?
Am'I misisng any step.
B. When we use the CM, do we have to dilute with regular medium in 30%, 50% ratio or use CM directly?