Cardiac progenitor cells are derived from mouse cardiac cells and sorted for Sca-1 and c-kit positive cells.
Adult mouse cardiac cells were depleted by collagenase (0.1%) for 30 min at 37°C, conditions lethal for most adult ventricular myocytes. Cells were further sorted by flow cytometry for stem cell markers including Sca-1 and c-kit. Cells were then labeled with Sca-1-biotin and incubated with antibiotin microbeads, purified by five to six cycles of magnetic selection.
Cells were grown in Medium-199 supplemented with FBS (10%), in 5% CO2, 37°C) on dishes coated with fibronectin (200 μg/ml).