Isolation and Culture of Rat Cardiac Fibroblasts

Cardiac fibroblasts were isolated from cardiac ventricles (combined left and right) of adult surplus rats (n = 31) from any age, weight, sex or breed. Most of the rats used were either from the Lewis or Wistar strain and aged between 5 and 52 weeks. Rat cardiac ventricular fibroblasts were isolated and cultured as previously described [9 (link),45 (link),46 (link)] in Dulbecco’s modified eagles medium (DMEM; no. 22320, Gibco, Invitrogen, Breda, the Netherlands) supplemented with 10% (v/v) fetal bovine serum (FBS, Gibco), gentamicin (50 µg/mL, Gibco), 1% (v/v) Insulin-Transferrin-Selenium-Sodium Pyruvate (ITS-A, Gibco) and basic fibroblast growth factor (1 ng/mL, Gibco) (“CF culture medium”). The vast majority of these cells are fibroblast-like cells and these primary fibroblasts were used between passage 1–3. Cardiomyocytes were isolated from the left ventricle of adult male Sprague Dawley rats (n = 6 age 10–20 weeks) essentially as described previously [32 (link),47 (link)]. Experiments were performed with approval of the Animal Ethical Committee of Maastricht University (DEC-2007-116, July 31, 2007) and conform to the national legislation for the protection of animals used for scientific purposes.