Name: Video: Mouse Liver Operation and Perfusion for Studying Hepatic Metabolism
Uploaded: 2023-10-06T15:00:00
Description: 500 Views. Mouse Liver Operation and Perfusion for Studying Hepatic Metabolism

mouse liver operation and perfusion for studying hepatic metabolism

In Situ Mouse Liver Perfusion Model to Study Metabolic Responses to Substrates

The liver is an essential organ in metabolism. It plays a key role in the control of whole-body energy balance by regulating glucose, lipid, and amino acid metabolism. The increase in liver diseases worldwide is emerging as a major global health burden, and more knowledge is needed about the pathophysiology and its consequences for liver functions.
Various in vitro models have been developed for research on the liver to complement in vivo studies. Isolated and cultured primary hepatocytes from rodents and humans are widely used. Non-parenchymal cells can be separated from hepatocytes using differential and gradient centrifugation, and the co-culture of different cell types is useful for studying intercellular crosstalk1. Although primary human hepatocytes are considered the golden standard for testing drug toxicity, several studies have shown that the hepatocytes rapidly dedifferentiate in tissue culture resulting in loss of hepatic functions2,3,4. Hepatocyte culture in a 3D spheroid system ameliorates the dedifferentiation, is more stable, and appears to mimic the liver in vivo to a higher degree than the traditional 2D culture systems5. Precision-cut liver slices are another well-established in vitro model that keeps the tissue architecture intact and contains the non-parenchymal cells present in the liver6. More advanced in vitro models include liver-on-a-chip7 and liver organoids8. However, with all these approaches, there is a loss of structural integrity and flow dynamics, including vectorial portal-hepatic vein flow, which likely impacts the generalizability.
The isolated perfused rat liver was first described by Claude Bernard in 18559, and is still used in various scientific fields for studies of liver biology, toxicology, and pathophysiology. Advantages of the perfused liver compared to the above-mentioned in vitro models include the maintenance of the hepatic architecture, the vascular flow, the hepatocyte polarity and zonation, and the interactions between hepatocytes and non-parenchymal cells. Compared to in vivo studies, the perfused liver allows the study of liver metabolism in an isolated manner avoiding extra-hepatic factors carried by the blood and with complete control over the experimental conditions. Several modifications have been made to improve the rat liver perfusion model over the years10,11,12,13. Although mice have been used for isolated perfused liver studies, less literature is available. Here, we present a method for in situ perfusion of the mouse liver by cannulation of the portal vein and the suprahepatic vena cava inferior to study the acute and direct metabolic responses to metabolic substrates and hormones as measured in the hepatic venous effluent from the mouse liver in real-time.

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Mouse Liver Operation and Perfusion for Studying Hepatic Metabolism   

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500 Views. Mouse Liver Operation and Perfusion for Studying Hepatic Metabolism

Video: Mouse Liver Operation and Perfusion for Studying Hepatic Metabolism   

The liver has numerous functions, including nutrient metabolism. In contrast to other in vitro and in vivo models of liver research, the isolated perfused liver allows the study of liver biology and metabolism in the whole liver with an intact hepatic architecture, separated from the influence of extra-hepatic factors. Liver perfusions were originally developed for rats, but the method has been adapted to mice as well. Here we describe a protocol for in situ perfusion of the mouse liver. The liver is perfused antegradely through the portal vein with oxygenated Krebs-Henseleit bicarbonate buffer, and the output is collected from the suprahepatic inferior vena cava with clamping of the infrahepatic inferior vena cava to close the circuit. Using this method, the direct hepatic effects of a test compound can be evaluated with a detailed time resolution. Liver function and viability are stable for at least 3 h, allowing the inclusion of internal controls in the same experiment. The experimental possibilities using this model are numerous and may infer insight into liver physiology and liver diseases.

Here, we present a robust method for in situ perfusion of the mouse liver to study the acute and direct regulation of liver metabolism without disturbing the hepatic architecture but in the absence of extra-hepatic factors.

The liver has numerous functions, including nutrient metabolism. In contrast to other in vitro and in vivo models of liver research, the isolated perfused ...