After hearts were rapidly washed in cool HEPES-Tyrode solutinn (in mmol/L: NaCl 143, KCl 5.4, NaH2PO4·2H2O 0.33, MgCl2·6H2O 0.5, CaCl2 1.8, glucose 5.5, and HEPES 5.0, pH 7.4 with NaOH), they were mounted to a Langendorff perfusion apparatus and perfused via the coronary circulation for 15 min with oxygenated Tyrode solution warmed to 37 ± 0.5°C.
Excised hearts were quickly transferred to a langendorf perfusion apparatus and perfused with calcium (Ca2+) free buffer containing (in mM); 90 NaCl, 10 KCl, 1.2 KH2PO4, 5 MgSO4.7H2O, 15 NaHCO30 30 taurine and 20 glucose for 5 min. The perfusion medium was then switched to Ca2+ free buffer containing collagenase (0.05%) and bovine serum albumin (0.2%).
The Langendorff-perfused heart was first used at the end of the 19th century, and comprises mounting the isolated heart on the aortic cannula of a Langendorff perfusion apparatus and retrogradely perfusing the heart at a constant flow or constant pressure, using blood or oxygenated buffer (typically Krebs–Henseleit solution).
Isolated rat hearts were immediately attached to the Langendorff perfusion apparatus (Phymep, France) and retrogradely perfused (Peristaltique pump, Gibson, Paris) at a constant perfusion pressure with oxygenated Krebs-Henseleit buffer containing NaCl (118 mM), KCl (4.7 mM), CaCl2 (1.9 mM), MgSO4 (1.2 mM), KH2PO4 (1.2 mM), NaHCO3 (25 mM), glucose (11 mM), pH7.4 [ 23].
The hearts were transferred to a perfusion apparatus (Hugo Sachs Elektronik Harvard Appartus GmbHbH, March-Hugstetten, Germany) and perfused in the Langendorff mode with modified Krebs-Henseleit solution (mM: 118 NaCl, 4.7 KCl, 1.75 CaCl2, 1.2 KH2PO4, 1.2 Mg24.9, 24.9 NaHCO3, 11.0 glucose) under continuously bubbling with carbogen (95%O2/5%5 % CO2) and a constant perfusion pressure of 80 mmHg.
Hearts from euthanized animals were mounted on a heart perfusion apparatus (Hugo Sachs Elektronik/Harvard Instruments, March-Hugstetten, Germany) and retrogradely perfused at 37 °C with oxygenated Krebs–Henseleit buffer at 80 mm Hg (constant pressure perfusion) as previously described [21].
The hearts were rapidly excised into ice-cold buffer, and mounted on a constant pressure (80 mmHg) Langendorff-perfusion apparatus and perfused with modified Krebs-Henseleit bicarbonate buffer in mmol/L: NaCl 118.5, NaHCO3 25.0, KCl 4.8, MgSO4 1.2, KH2PO4 1.2, CaCl2 1.7 and glucose 5.0 or 11.0.
The hearts were removed and mounted on a Langerdorff-perfusion apparatus.
Hearts were rapidly excised and placed in ice-cold buffer and mounted on a constant pressure (100 mmHg) Langendorff-perfusion apparatus.
The aorta was cannulated and the heart was perfused in a Langerdorf-type apparatus with perfusion buffer containing (in mmol/l) NaCl 120.4, KCl 14.7, KH2PO4 0.6, Na2HPO4 0.6, MgSO4 1.2, NaHCO3 4.6, taurine 30, 2,3-butanedione monoxime 10, glucose 5.5, and HEPES 10 (pH 7.0), for 4 min at a flow rate of 4 ml/min.
To determine if the EDCs were present in the myocardial interstitium, blood was flushed out of the hearts by retrograde perfusion on a Langendorff apparatus for approximately three minutes at room temperature.
