My study is designed to treat the confluent human smooth muscle cells with clinical human plasma and look for calcification in the cells. For the induction of calcification, I am using ossification medium (OM) containing 1.58 mM/L calcium gluconate, 0.1 μM/L

dexamethasone, 0.5 mM/L ascorbic acid and 2.5 mM/L beta glycerol phosphate. After adding the OM, the adherent cells are making a gel like structure getting detached leaving a few cells attached to the plate. I need a really detailed protocol to follow.