A CDAA-HFAT (Research Diets, New Brunswick, NJ) or choline-supplemented, L-amino acid-defined control diet was fed to mice for 10 weeks starting at the age of 8 weeks. As a second dietary NASH model, a Western diet (high fat, fructose and cholesterol diet and water supplemented with sucrose/fructose) (AIN-76A Western Diet, TestDiet, St. Louis, MO) or chow control diet was fed to mice for 20 weeks starting at the age of 8 weeks. For dietary experiments, groups (knock-outs and controls) were cohoused. Mice fed CDAA diet show severe NASH characterized by steatohepatitis and fibrosis.
Ain 76a western diet
Manufactured by TestDiet
Sourced in Canada
AIN-76A Western Diet is a nutritionally complete rodent diet formulated to induce obesity and metabolic disorders. It contains increased levels of fat and carbohydrates compared to a standard rodent chow. This diet is commonly used in research studies focused on diet-induced obesity and associated conditions.
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Lab products found in correlation
Cdaa hfat, by Research Diets (1 mentions)
Male sprague dawley rat, by Charles River Laboratories (1 mentions)
C57bl 6ncrl, by Charles River Laboratories (1 mentions)
Lna scr, by Qiagen (1 mentions)
Albumin cre alb cre mice, by Jackson ImmunoResearch (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
7 protocols using ain 76a western diet
1
Dietary NASH Induction in Mice
2
Sprague-Dawley Rat High-Fat Diet Protocol
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All of the experiments were conducted following the guidelines of the Canadian Council of Animal Care and were approved by Memorial University's Institutional Animal Care Committee. Male Sprague–Dawley rats (Charles River, Saint Constant, Canada or Memorial University breeding colony) and C57BL/6NCrl (Charles River, Saint Constant, Canada) were fed ad libitum with a standard chow (LabDiet autoclavable rodent diet 5010, 12.7% fat) or a palatable HFD (TestDiet AIN-76A Western Diet, 40.1% fat). Body weight and food intake were measured weekly. Animals were 3 weeks old at the start of the feeding period unless stated otherwise. For 1-day HFD, the feeding period was staggered so that the rats were 4 weeks old at the time of electrophysiological recordings.
3
High-fat, high-cholesterol diet in mice
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Ten-week-old male C57BL/6J were housed 5 mice per cage, and cages were randomized to a diet for 24 weeks: standard chow (Diet Pico Laboratory Rodent Diet) or FFC diet, which included a high-fat and -cholesterol chow (AIN-76A Western Diet; 1810060; Test Diet)and drinking water that was supplemented with fructose (23.1 g/L, MilliporeSigma, F2543) and glucose (18.9 g/L,MilliporeSigma, 49158) as previously described (49 (link)).
4
High-Fat, High-Cholesterol, High-Fructose Diet in Mice
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The HFCF diet contained 40% fat, 0.2% cholesterol (AIN‐76A Western diet from TestDiet), and 4.2% fructose (in drinking water). Eight‐week‐old male mice were used and fed an HFCF diet for 16 or 20 weeks. The high‐fat diet (HFD) containing 60% kcal from fat was purchased from Research Diets (Cat. #D12492). Eight‐week‐old mice were used and fed an HFD for 20 weeks.
5
Rip1, Rip3 Kinase Mutant Mice on High-Fat Diet
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All procedures using animals were approved by the Cleveland Clinic Institutional Animal Care and Use Committee. Rip1K45A/K45A and Rip3K51A/K51A mice were originally generated by GlaxoSmithKline. Briefly, Rip1K45A/K45A and Rip3K51A/K51A mice were generated through homologous recombination, utilizing targeting construct that mutated the catalytic lysine residue to alanine (K45A and K51A, respectively), resulting in the complete elimination of kinase activity (17 (link), 18 (link)). Both strains are viable and fertile backcrossed to a C57BL/6J background and WT controls were wild-type littermates.
Male mice (5-6 weeks of age) were allowed free access to a chow diet (Chow) containing 6% fat/13.0 kJ/g (#2918, Teklad Mills, Madison, WI) or FFC diet (AIN-76A Western diet, #5342, TestDiet, St. Louis, MO) for 12 weeks. The FFC diet contained 40% energy as fat (12% saturated fatty acid, 0.2% cholesterol) with fructose and glucose added to the water (42 g/L final concentration).
Food intake was similar between genotypes in response to FFC diet. As expected, body and liver weights were higher with FFC feeding in all genotypes compared to chow-fed mice. Rip1K45A/K45A and Rip3K51A/K51A mice gained 12% and 11% less, respectively, on the FFC diet compared to WT (Supplementary Figure S1
Male mice (5-6 weeks of age) were allowed free access to a chow diet (Chow) containing 6% fat/13.0 kJ/g (#2918, Teklad Mills, Madison, WI) or FFC diet (AIN-76A Western diet, #5342, TestDiet, St. Louis, MO) for 12 weeks. The FFC diet contained 40% energy as fat (12% saturated fatty acid, 0.2% cholesterol) with fructose and glucose added to the water (42 g/L final concentration).
Food intake was similar between genotypes in response to FFC diet. As expected, body and liver weights were higher with FFC feeding in all genotypes compared to chow-fed mice. Rip1K45A/K45A and Rip3K51A/K51A mice gained 12% and 11% less, respectively, on the FFC diet compared to WT (
6
Hepatocyte-specific miR-34a Knockout Mice
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miR-34afl/fl mice, albumin-Cre (Alb-Cre) mice, and C57BL/6J mice were purchased from the Jackson Laboratory (Bar Harbor, Maine, USA) [4 (link)]. miR-34afl/fl mice were crossed with Alb-Cre mice to generate germline hepatocyte-specific miR-34a−/− (miR-34agHep−/−) mice and control littermates (miR-34afl/fl mice). AAV8-TBG-Null or AAV8-TBG-Cre was i.v. injected into the miR-34afl/fl mice to generate control mice (miR-34afl/fl mice) or adult-onset hepatocyte-specific miR-34a−/− (miR-34aHep−/−) mice, respectively. Locked nucleic acids (LNA) against scramble sequences (LNA-Scr) or miR-34a (LNA-miR-34a) were synthesized by Qiagen and i.p. injected into the mice at a dosage of 10 mg/kg/week. All of the mice were housed in a temperature- and humidity-controlled room with a 12-h light/12-h dark cycle under pathogen-free conditions. The high-fat/cholesterol/fructose (HFCF) diet contained 40% fat/0.2% cholesterol (AIN-76A Western diet from TestDiet) and 4.2% fructose (in drinking water). Unless otherwise stated, 2-month-old male mice were used and fed an HFCF diet for 16 weeks. The mice were fasted for 5–6 h during the light cycle prior to euthanasia. All the of animal experiments were approved by the Institutional Animal Care and Use Committee at Northeast Ohio Medical University (NEOMED).
7
High-Fat Diet Induces Metabolic Dysfunction
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Ten-week-old male C57BL/J6 were housed 5 mice per cage, and cages were randomized to a diet for 24 weeks: standard chow (Diet Pico Laboratory Rodent Diet) or high fat, fructose, and cholesterol (FFC) diet which included a high fat and cholesterol chow (AIN-76A Western Diet; 1810060; Test Diet) and drinking water supplemented with fructose (23.1 g/L, Sigma F2543) and glucose (18.9 g/L, Sigma 49158), as previously described (23 (link)).
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