Objective: Examine the effects of metformin supplementation on lifespan and survival in B6C3F1 mice through long-term dietary intervention
This is a Metformin Longevity Study - B6C3F1 protocol using mouse as the model organism. The procedure involves 7 procedural steps, 4 equipment items, 3 materials. Extracted from a 2013 paper published in Nature Communications.
Model and subjects
mouse • B6C3F1 • male • 12 months at treatment start • not specified • 333
Study window
Estimated timing pending
Core workflow
Animal Acquisition and Baseline Housing • Diet Preparation and Metformin Incorporation • Diet Storage
Primary readouts
Key equipment and reagents
Verified items
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Direct vendor links
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Male B6C3F1 mice obtained from Harlan Breeders (Indianapolis) and maintained on ad libitum chow diet (Diet #5001, Purina Mills, Richmond, IN) prior to treatment assignment
Note: Mice were randomly assigned to treatment groups at 12 months of age
“male mice (Harlan Breeders; Indianapolis) were randomly assigned to treatment groups at 12 months of age”
Metformin from Caraco Pharmaceutical Laboratories mixed with powdered AIN-93M diet at 100 mg/kg diet (0.1% w/w) and cold-pressed into 1-gram pellets by BioServ. Chow produced every 3 months, never exceeding 50°C during processing
Note: Processing temperature controlled to ensure metformin stability. Light exposure minimized during production and storage
“Metformin was mixed with powdered diet and cold-pressed into 1-g pellets by BioServ. The chow was produced every 3 months during the length of the study, never was permitted to exceed 50 °C”
Stock metformin and all chow stored in darkness at 4°C to maintain metformin stability. Light/dark cycle in mouse facility not altered
Note: Storage conditions maintained to prevent metformin degradation
“Stock metformin and all chow were stored in the dark at 4 °C. The light/dark cycle in the mouse facility was not altered”
At 12 months of age, mice randomly assigned to control group (shifted to daily feeding with 13.3 kcal/day/mouse of AIN-93M control diet) or metformin group (shifted to identical quantity of AIN-93M diet supplemented with metformin at 100 mg/kg diet, approximately 10.6 mg/kg body weight per day)
Note: All mice fed daily. With rare exceptions, all food was consumed each day
“male mice (Harlan Breeders; Indianapolis) were randomly assigned to treatment groups at 12 months of age. In the control group, mice were shifted from ad lib chow feeding to daily feeding with 13.3 kcal/day/mouse of control diet”
Food intake and body weight measured on biweekly or bimonthly basis throughout the duration of the study
Note: Measurements continued until animal death or euthanasia
“Food intake and body weight were measured on a biweekly or bimonthly basis for the duration of the study”
Mice monitored for survival throughout study. Euthanasia criteria based on independent veterinary assessment according to AAALAC guidelines, only when animal condition incompatible with continued survival. Every animal found dead or euthanized was necropsied for pathology score
Note: Veterinary assessment required for euthanasia decisions. All animals underwent necropsy
“The criteria for euthanasia was based on an independent assessment by a veterinarian, according to AAALAC guidelines and only cases, where the condition of the animal was considered incompatible with continued survival, are represented in the curves. Every animal found dead or euthanized was necropsied for pathology score”
Survival curves plotted using Kaplan-Meier method, which includes all available animals at each time point
Note: Statistical analysis method for survival data
“Survival curves were plotted using the Kaplan-Meier method, which includes all available animals at each time point”
This section explains what the experiment is doing, which readouts matter, what the data artifacts usually look like, and how the analysis should flow from raw capture to reported result.
Examine the effects of metformin supplementation on lifespan and survival in B6C3F1 mice through long-term dietary intervention
Objective
Examine the effects of metformin supplementation on lifespan and survival in B6C3F1 mice through long-term dietary intervention
Subjects
From papermouse • B6C3F1 • male • 12 months at treatment start • not specified
Sample count
From paper333
Cohort notes
From paperMale mice randomly assigned to treatment groups at 12 months of age.
Animal Acquisition and Baseline Housing (until 12 months of age)
Diet Preparation and Metformin Incorporation (every 3 months throughout study)
Diet Storage (throughout study duration)
Treatment Group Assignment and Diet Transition (at 12 months of age, continuing for remainder of life)
Lifespan and survival curves
From paperSurvival curves were plotted using the Kaplan-Meier method, which includes all available animals at each time point
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Body weight changes
From paperSurvival curves were plotted using the Kaplan-Meier method, which includes all available animals at each time point
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Food intake
From paperSurvival curves were plotted using the Kaplan-Meier method, which includes all available animals at each time point
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Pathology scores from necropsy
From paperSurvival curves were plotted using the Kaplan-Meier method, which includes all available animals at each time point
Artifact type
Endpoint measurements summarized by group or timepoint
Comparison focus
Compare endpoint magnitude between groups, timepoints, or both
Lifespan and survival curves
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Body weight changes
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Food intake
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Pathology scores from necropsy
From paperRaw artifact
Per-sample or per-animal endpoint measurements collected during the experiment
Processed artifact
Structured table with cleaned measurements ready for comparison
Final reported form
Summary statistics and between-group or across-timepoint comparisons
Acquisition
Collect raw experimental outputs with enough metadata to preserve sample identity, condition, and timing.
Preprocessing / cleaning
Survival curves were plotted using the Kaplan-Meier method, which includes all available animals at each time point
Scoring or quantification
Quantify the primary readouts for this experiment: Lifespan and survival curves; Body weight changes; Food intake; Pathology scores from necropsy.
Statistical comparison
Statistical method not yet structured for this page.
Reporting output
Report representative outputs alongside summary comparisons for Lifespan and survival curves, Body weight changes, Food intake, Pathology scores from necropsy.
Source links and direct wording from the methods section for validation and deeper review.
Citation
Alejandro Martin-Montalvo et al. (2013). Metformin improves healthspan and lifespan in mice. Nature Communications
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Direct vendor pages are linked from the protocol above. This section stays focused on the full comparison view and the prep checklist.
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BioServ • not specified • not specified • not mentioned
not specified • not specified • not specified • not mentioned
not specified • not specified • not specified • not mentioned
not specified • not specified • not specified • not mentioned
Caraco Pharmaceutical Laboratories • not applicable • not specified • not mentioned
BioServ • not applicable • Diet No. F05312 • not mentioned
Purina Mills • not applicable • Diet # 5001 • not mentioned
not applicable - statistical method • not mentioned
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Current status surfaces were computed from experiment data updated Feb 28, 2026.
Source access
Jump back into the original paper or the methods evidence section when you need exact wording, exclusions, or method-specific caveats.
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Steps
7
Evidence Quotes
14
Protocol Items
7
Linked Products
0
Canonical Sync
Pending
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Steps
7
Evidence
14
Specific Products
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Canonical Sync
Pending
What this score means
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Computed from the current experiment record updated Feb 28, 2026.
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