Mesenchymal stem cells derived from placental amnions after treating the tissue with pancreatin and collagenase.
The placental amnions is cut into small pieces and digested with pancreatin (2.5 g/L) for 30 min at 37◦C. After terminating the digestion with FCS, the tissue is further digested with collagenase (1 g/L) for 60 min and then filtered through a stainless steel mesh to form a single cell suspension. Cells are collected after centrifugation and plated in culture flasks in DMEM/F12 supplemented with FCS (10%) and bFGF (20 g/L) and grown at 37◦C, 5% CO2. Medium is replaced every 3–4 days.