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Essential Insights | Why Does My Dio Model-Making Always Fail?

Introduction

The Dio mouse model is easy to establish and widely applicable, leading many researchers to undertake modeling themselves. However, if the operator lacks sufficient experience or fails to attend to critical details, the mice may exhibit slow weight gain or even weight loss, which can subsequently impair glucose metabolism.

Today, drawing on relevant reports and real-world challenges, we’ve compiled and analyzed key insights to share practical experience and tips for constructing Dio‑model mice, helping researchers enhance the stability and reproducibility of their Dio models.

 

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High-fat‑induced obese mice are a commonly used model in current research on obesity and type 2 diabetes. Typically, C57BL/6J mice around 6 weeks of age are selected, and a high‑fat diet providing 60 kcal% of energy is administered to establish the model. The phenotypes observed in these mice closely resemble those seen in patients with early‑stage diabetes and metabolic disorders, including increased body weight, elevated blood glucose levels, and impaired glucose tolerance.

 
Feed

Dio mice are typically fed a 60% high-fat diet—D12492. Currently, this diet is available in two main forms: imported (Research Diets) and domestically produced. In practice, the two types show little difference in performance and both can meet the requirements for model establishment. Researchers may choose based on their budget.

Due to the high fat content of the diet, it should be stored temporarily refrigerated (4°C) or frozen (-20°C). Replace the food 1–2 times per week to prevent spoilage. Many studies report that high-fat diets do not require additional thawing; however, based on caretakers’ observations, mice prefer the diet when it is brought to room temperature prior to feeding. Place a generous amount of feed in the cage to ensure that mice can access it at any time. This practice helps promote weight gain, particularly when the mice become large and struggle to maintain an upright posture, as they find it easier to eat within the cage.

Many experimental protocols require measuring food intake in mice, but the feed at the bottom of the cage can compromise the accuracy of these measurements. Therefore, we believe that food intake is only one aspect of the data; the primary goal remains ensuring that mice can readily access an adequate supply of food.

 

 

Drinking water

Ensure that mice have constant access to water. Many researchers focus primarily on the diet when establishing experimental models, often neglecting adequate hydration. Regularly monitor the water level in the drinking bottles; high‑fat‑diet–fed mice typically consume more water than standard‑diet–fed mice. Maintaining proper hydration in high‑fat‑diet–fed mice can also help promote weight gain.

 

 

Body weight

Data indicate that the initial body weight of mice also influences subsequent weight gain. Therefore, when purchasing mice, it is recommended to select individuals with larger, uniform body weights—typically around 18 g at 6 weeks of age—and begin feeding them at this stage.

During transportation, a reduction in body weight is normal for mice. Body weight typically returns to baseline within one to two weeks after arrival. Therefore, mice should be provided with an acclimatization period of 1–2 weeks prior to the start of the experiment.

Click here to view the mouse body weight change table.

 

 
Environment

Providing mice with a comfortable living environment also promotes weight gain. Littermates are less likely to fight. Mice housed individually may be less prone to obesity than those kept in groups; however, if each cage contains too few animals, they may expend more calories on thermoregulation. Aside from routine cage changes and the replacement of food and water, minimize disturbances to the mice as much as possible.

Do not house mice from different cages or packaging boxes together. Mixed-group housing can lead to aggression, which may impair weight gain.

Temperature control is also an important husbandry factor: it is recommended to house high-fat‑diet mice at a constant temperature of approximately 26°C. Excessively high housing temperatures can lead to heat‑induced mortality, whereas keeping these mice in a standard animal facility at 22±2°C requires them to expend energy to maintain their body temperature.

However, some studies suggest that the optimal housing temperature for high-fat‑diet mice is 29–32°C. Please monitor your mice closely; if their fur becomes damp, it indicates that the room temperature is too high, and you should adjust it promptly.

 

 

Bedding material

Many animal facilities commonly use corn cob bedding, which has little impact on standard mouse strains. However, in high‑fat–induced mouse models, the poor palatability of the high‑fat diet often leads mice to gnaw on the corn cob bedding and consume it voraciously, thereby reducing their intake of the high‑fat diet and impairing weight gain. Consequently, when establishing Dio‑type models, we recommend using wood shavings as bedding. Additionally, incorporating shredded paper can help maintain a stable body temperature for obese mice.

 

 

Blood glucose testing

The optimal time to measure blood glucose levels in high-fat‑diet–fed mice is early morning. Since mice feed at night, their morning glucose readings reflect non‑fasting levels. To ensure consistency in research data, it is crucial to sample mice at the same time of day when assessing their blood glucose concentrations.

 

 

Backup mice

There is a certain failure rate in generating the Dio mouse model; therefore, we recommend ordering an additional 5–10% of mice for each experimental batch, as some mice may be insensitive to high-fat diets.

 

Reference link:

https://www.jax.org/news-and-insights/jax-blog/2014/may/seven-tips-every-Dio-mice-user-should-know

Seeley, R.J., MacDougald, O.A. Mice as experimental models for human physiology: when several degrees in housing temperature matter. Nat Metab (2021).

https://doi.org/10.1038/s42255-021-00372-0

 

Aniphe BioLab can provide you with: DIO mouse model , Standard in-stock items: 12–18 weeks; feel free to inquire.

 

 

 

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