Liver perfusion experiments, the closest in vitro research system to the in vivo state in drug liver metabolism studies, retains the basis of intact cellular and tissue structure, allows for the separate examination of hepatobiliary disposition of chemical xenobiotics, and can greatly improve the efficiency of drug development. The methods used for liver perfusion experiments rely primarily on perfusion devices.
Principle
The basic principle of hepatic perfusion is to surgically circulate the liver under anesthesia. A peristaltic pump pumps a Krebs-Henseleit or Krebs-Ringer solution containing low molecular weight glucose saturated with oxygen and carbon dioxide (955) gases at a constant rate into the circulatory tubing, which flows through a filtration device, a warming device, a portal cannula, the liver, and finally out of the superior or inferior vena cava. It flows through a filtration unit, a warming unit, a portal vein cannula, the liver, and finally out of the superior or inferior vena cava, where the effluent can be sampled and analyzed or returned to the reservoir for recirculation, allowing the liver to maintain its normal physiological and biochemical functions for a period of time.
Appliance
Hepatic perfusion assays can be used to dynamically study the metabolic changes of chemical xenobiotics in the liver and their effects on liver function.
Operation method
liver perfusion assay
Principle
The basic principle of hepatic perfusion is to surgically circulate the liver under anesthesia. A peristaltic pump pumps a Krebs-Henseleit or Krebs-Ringer solution containing low molecular weight glucose saturated with oxygen and carbon dioxide (955) gases at a constant rate into the circulatory tubing, which flows through a filtration device, a warming device, a portal cannula, the liver, and finally out of the superior or inferior vena cava. It flows through a filtration unit, a warming unit, a portal vein cannula, the liver, and finally out of the superior or inferior vena cava, where the effluent can be sampled and analyzed or returned to the reservoir for recirculation, allowing the liver to maintain its normal physiological and biochemical functions for a period of time.
Materials and Instruments
Equipment: perfusion device; thermostatic circulation system, perfusate circulation system and gas exchange system; surgical instruments. Move The liver perfusion experiment was performed as follows: For more product details, please visit Aladdin Scientific website.
Experimental animals: 200-250 g rats
A, After the rat was anesthetized by intraperitoneal injection of sodium pentobarbital 50 mg/kg, it was placed supine on the operating table.
B, The abdominal cavity was U-shaped clipped open, and the abdominal organs were shifted to the left side of the torso, exposing the bile ducts and the hepatic portal vein.
C, Separation of the bile ducts, insertion of the bile duct conduit, and fixation.
D, Separation of the portal vein, ligating of the hepato-splenic branches, and threading a surgical wire and tie a live sleeve, make portal vein cannula, quickly fix it and start perfusion.
E, cut the abdominal segment of the inferior vena cava, quickly open the thoracic cavity, cut the thoracic segment of the inferior vena cava on the side against the diaphragm, and flush out the residual blood in the liver.
F, separate out the liver intact and undamaged, and transfer it to the organ tray of the perfusion apparatus or put it in the original position.
G, flush out the residual blood in the liver with the perfusion media that is pre-warmed at 37 ℃.
H, flush out the residual blood in the liver with a perfusion media that doesn't contain erythrocyte H. Equilibrate the liver with perfusion medium without erythrocytes and other additives and the subject drug for 10-20 minutes, then switch to perfusion medium containing the subject drug and other additives and adjust the perfusion rate.