We obtained CFs and primary CMs from the hearts of 1-3-day-old Sprague-Dawley (SD) rats by trypsin digestion. After washing with ice-cold PBS, the heart was cut into 1 mm3 pieces and digested with 0.125% trypsin. The digestion was repeated 6-7 times, and the cells were thoroughly mixed and plated in a sterile 10 cm culture dish. After 2 h of cell attachment, the attached CFs and CMs from the medium were collected. The adherent CFs or CMs were cultured in DMEM/F12 containing 10% FBS and 1% antibiotic for 24 h, and then the medium was changed every 48 h.

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