The porcine intestinal mucosa require large amounts of energy for nutrient processing and cellular
functions and is vulnerable to injury by weaning stress involving bioenergetics failure. The mitochondrial
bioenergetic measurement in porcine enterocytes have not been defined. The present study was to
establish a method to measure mitochondrial respiratory function and profile mitochondrial function of
IPEC-J2 using cell mito stress test and glycolysis stress test assay by XF24 extracellularflux analyzer. The
optimal seeding density and concentrations of the injection compounds were determined to be 40,000
cells/well as well as 0.5mM oligomycin, 1mM carbonyl cyanide p-trifluoromethoxy-phenylhydrazone
(FCCP) and 1 mM rotenone & antimycin A, respectively. The profiles of mitochondrial respiration and
glycolysis confirmed that porcine enterocyte preferentially derived much more energy from glutamine
than glucose. These results will provide a basis for further study of mitochondrial function and bioenergetics of the porcine small intestine.