XFe96 Analyzer Power Pak

At the department of experimental hepatology, we have long focused on toxic liver injury, including mechanisms of action of model hepatotoxins and substances with potential hepatoprotective effects, hepatic regeneration induced by partial hepatectomy or toxic liver injury, sensitivity of the liver to hepatotoxic agents and regenerative response in the field of liver affected by non-alcoholic fatty liver disease (NAFLD, NASH), extrahepatic cholestasis induced by ligation d. choledochus, and the possibility of influencing the development of subsequent liver injury.

The liver is an important organ of the body, it performs, among other things, a number of irreplaceable functions in intermediary and energy metabolism, it is the main site of biotransformation and elimination of xenobiotics and many substances of endogenous origin. The importance of proper assessment of mitochondrial function is emphasized by findings that document that impaired hepatic mitochondrial function accompanies most common metabolic, nutritional, and other diseases of civilization. Mitochondrial function can be studied in a variety of models including liver homogenate, mitochondria and hepatocytes isolated from the liver under in vivo conditions; isolated hepatocytes, stabilized cell lines or tumor cell lines can be used in in vitro experiments. For the assessment of mitochondrial function, it is essential to have instrumentation and standard operating procedures harmonised with other laboratories to allow comparison of results obtained at other sites.

In our laboratory, we have so far used high-performance respirometry using the Oroboros-2k Oxygraph. By appropriately combining different substrates, extenders and specific inhibitors, the oxygraph allows the assessment of different biotransformation energy pathways, individual respiratory complexes and functions (respiratory control index, ATP production rate, integrity of the outer and inner mitochondrial membrane). Tissue homogenates, intact or permeabilized cells, isolated mitochondria and permeabilized tissues can be analyzed. The disadvantage is that this instrument allows simultaneous analysis of only 2 samples in one oxygraph and it is not possible to analyze cells adhered to the surface.

The Core Facilities project allowed us to acquire a unique instrument for cellular metabolic analysis - the Agilent Seahorse Bioscience XFe96 analyzer.