Source Paper
Elise F. Hoek-van den Hil, Evert M. van Schothorst, Inge van der Stelt, Hans J. M. Swarts, Marjanne van Vliet et al.
Genes & Nutrition • 2015
Objective: Weekly monitoring of body weight and food intake throughout a 12-week dietary intervention period in mice receiving normal-fat or high-fat diets with or without flavonoid supplementation
Gather these items before starting the experiment. Check off items as you prepare.
Harlan Laboratories • Not specified • Not specified • Not mentioned
Research Diets Services B.V. • Not specified • Not specified • Not mentioned
Research Diets Services B.V. • Not specified • Not specified • Not mentioned
Sigma • Not specified • Not specified • Not mentioned
Bioconnect • Not specified • Not specified • Not mentioned
Sigma • Not specified • Not specified • Not mentioned
Fuzhou Corona Science & Technology Development Co., Ltd. • Not specified • Not specified • Not mentioned
Medox, Polyphenols Laboratories • Not specified • Not specified • Not mentioned
Greiner Bio-one • Not specified • Not specified • Not mentioned
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Eighty-four male C57BL/6JOlaHsd mice arrived at 9 weeks of age and were individually housed under controlled conditions
Note: Controlled conditions: 12h/12h light-dark cycle, 55±15% humidity, ad libitum access to food and water
“Eighty-four male C57BL/6JOlaHsd mice (Harlan Laboratories, Horst, The Netherlands) were individually housed under controlled conditions (temperature 21°C, 12 h/12 h light–dark cycle, 55 ± 15 % humidity), with ad libitum access to food and water. At arrival, the mice were 9 weeks of age.”
During the first 5 days of the 3-week adaptation period, mice were fed standard Harlan chow diet
Note: Ad libitum access
“During the first 5 days of a 3-week adaptation period, mice were fed a standard Harlan chow diet”
Following the initial 5-day period, mice were fed a standardised semi-synthetic normal-fat diet (NF, 10 energy% fat) with the same dietary constituents as the intervention high-fat diet
Note: Ad libitum access. This diet has the same constituents as the HF diet but with carbohydrates substituted with fats in the HF diet
“followed by a standardised semi-synthetic normal-fat diet [NF, 10 energy% (en%) fat] with the same dietary constituents as the intervention high-fat diet (HF, 40 en%) in which carbohydrates were substituted with fats”
At the start of the 12-week intervention period, mice were stratified based on body weight over 7 groups (n=12 per group) to obtain identical groups for this important parameter
Note: Stratification ensures balanced groups for the intervention
“At the start of the 12-week intervention period, mice were stratified based on body weight over 7 groups ( n = 12)”
One group of mice continued on normal-fat diet (NF), while the other six groups received high-fat diet (HF) with or without supplementation of different flavonoids (HF+flavonoids). Flavonoids were added in equimolar amounts to HF (0.01 mol/kg diet): quercetin 0.33% (w/w), hesperetin 0.33%, epicatechin 0.32%, apigenin 0.29%, and anthocyanins 0.5%
Note: Ad libitum access to assigned diet. Flavonoid amounts based on previous results showing effectiveness of quercetin at this concentration
“One group of mice continued on NF, while the other six groups of mice received HF with or without supplementation of different flavonoids (HF + flavonoids). Flavonoids were added in equimolar amounts to HF (0.01 mol/kg diet)”
Body weight was measured weekly throughout the 12-week intervention period
Note: Monitoring continues throughout the entire intervention period
“Body weight and food intake were monitored weekly”
Food intake was measured weekly throughout the 12-week intervention period
Note: Monitoring continues throughout the entire intervention period
“Body weight and food intake were monitored weekly”
Faeces were collected from mice during weeks 11 and 12 of the intervention period
Note: Collection during final 2 weeks of intervention
“Faeces were collected in weeks 11 and 12”
At the end of the intervention, all mice were fasted for 2-4 hours during the light phase
Note: Fasting conducted during light phase of light-dark cycle
“At the end of the intervention, all mice were fasted for 2–4 h during the light phase”
Mice were anesthetised by inhalation of 5% isoflurane using O2 as a carrier
Note: Conducted after fasting period
“anesthetised by inhalation of 5 % isoflurane using O 2 as a carrier”
Blood was sampled via orbital extraction into serum collection tubes (Greiner Bio-one, Longwood, USA)
Note: Serum was separated and stored at -80°C
“Blood was sampled via orbital extraction in collect serum tubes (Greiner Bio-one, Longwood, USA) and stored at −80 °C after obtaining serum”
After blood collection, mice were killed by cervical dislocation. Liver, epididymal white adipose tissue (epiWAT), and mesenteric white adipose tissue (mesWAT) were dissected, weighed, and snap frozen in liquid nitrogen
Note: Tissues were stored at -80°C after snap freezing
“After blood collection, mice were killed by cervical dislocation, and liver, epididymal and mesenteric white adipose tissues (epiWAT and mesWAT, resp.) were dissected, weighted and snap frozen in liquid nitrogen and stored at −80 °C”
Individually housed under controlled conditions (temperature 21°C, 12h/12h light-dark cycle, 55±15% humidity), with ad libitum access to food and water. One HF+quercetin mouse excluded from analyses due to nasal abscess, resulting in n=83 for analysis.