A modified choline-deficient, ethionine-supplemented diet reduces morbidity and retains a liver progenitor cell response

The choline-deficient, ethionine-supplemented (CDE) dietary model induces chronic liver damage, and stimulates liver progenitor cell (LPC) mediated repair. Long-term CDE administration leads to hepatocellular carcinoma in rodents and lineage tracing studies show LPCs differentiate into functional hepatocytes in this model. The CDE diet was first modified for mice by our laboratory by separately administering choline-deficient chow and ethionine in the drinking water. Whilst this CD+E diet is widely used, concerns with variability in weight loss, morbidity, mortality, and LPC response have been raised by researchers who have adopted this model. We propose these inconsistencies are due to differential consumption of chow and ethionine in the drinking water, and combining the ethionine into the choline-deficient diet and altering the strength, will achieve better outcomes. Methods: C57Bl/6 mice, 4 and 5 weeks of age, were fed an all-inclusive CDE diet of various strengths (67% to 100%) for three weeks. The LPC response was quantitated and cells lines were derived. Results: Animal survival, LPC response, and liver damage are correlated with CDE diet strength. The 67% and 75% CDE diet administered to mice older than 5 weeks and greater than 18g provides a consistent and acceptable level of animal welfare and induces a substantial LPC response, permitting their isolation and establishment of cell lines. This study shows that an all-inclusive CDE diet for mice reproducibly induces an LPC response conducive to in vivo studies and isolation, whilst minimizing morbidity and mortality. D is ea se M o de ls & M ec ha ni sm s • D M M • A dv an ce a rt ic le LIST OF ABBREVIATIONS ALT (alanine aminotransferase), CDE (choline-deficient, ethionine-supplemented), CD+E (choline-deficient chow with ethionine-supplemented water), GGT (γ-glutamyltransferase) HCC (hepatocellular carcinoma), HNF4α (hepatocyte nuclear factor 4 alpha), LPC (liver progenitor cell), BMOL3 (bipotential murine oval liver 3), PanCK (pan cytokeratin), PBS (phosphate-buffered saline), RT (room temperature), WEM (Williams’ E medium) D is ea se M o de ls & M ec ha ni sm s • D M M • A dv an ce a rt ic le