Insulin transferrin selenium sodium pyruvate its a

Omentum-derived MCs were obtained from nonuremic patients undergoing abdominal surgery; the omentum samples were digested in 0.05% trypsin and 0.02% EDTA to isolate MCs [14 (link), 29 (link)]. All MCs isolated from the omentum were then incubated in culture medium consisting of Earle M199 medium (Gibco, NY, USA) supplemented with 10% fetal calf serum (Biological Industries, Kibbutz Beit-Haemek, Israel), insulin-transferrin-selenium-sodium pyruvate (ITS-A) (Gibco), 100 μg/mL of streptomycin (MDBio, Inc., Taipei, Taiwan), 63.6 μg/mL of penicillin G (Sigma-Aldrich, MO, USA), and 250 ng/mL Fungizone (MDBio, Inc.).

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.