Quality Evaluation Index of Oxidative Stress Cell Model
Creative Bioarray has been committed to providing customers with high-quality services. Therefore, we have established a set of mature judgment indicators of oxidative stress cell model to evaluate the quality of the model.
In order to successfully establish the oxidative stress cell model, after the cells are treated by the stressor, it is necessary to detect the cell damage, evaluate whether the stressor causes the oxidative damage of the cells, and the degree of damage, so as to select the best modeling conditions.
Generally speaking, we mainly evaluate by synthesizing the test results of the following four aspects.
- Cell viability
- Intracellular ROS level
- lipid peroxide
- Antioxidant enzymes
Cell Viability Test
We generally use MTT method and CCK-8 method to evaluate the cell survival rate.
The accuracy and accuracy of CCK-8 method are higher than that of MTT method, and this method is simple and rapid, with little harm to the environment. Therefore, when constructing the oxidative stress cell model, we give priority to CCK-8 method to detect the cell survival rate.
Evaluation of Total Intracellular ROS Level
We usually use the dichlorofluorescein diacetate probe (dcfda) method, which is the most direct index to evaluate the degree of cellular oxidative stress.
Evaluation of Lipid Peroxidation Products
Lipid peroxidation products can cause cell damage by attacking unsaturated fatty acids on cell membrane, so they can directly reflect oxidative damage. Malondialdehyde (MDA) is a common index in cell model, which is detected by thiobarbituric acid (TBARS).
Antioxidant enzymes Detection
Antioxidant related enzymes can indirectly reflect the antioxidant response in cells, such as glutathione peroxidase (GSH PX), catalase (CAT), superoxide dismutase (SOD), etc. they can convert free radicals into non-toxic forms. They are an important part of the intracellular antioxidant defense system and affect the dynamic balance between oxidation and antioxidation in vivo, The increase or decrease of the number of these enzymes can reflect the oxidation state in cells, so the activity of antioxidant enzymes is often used as the detection index when constructing the cell model of oxidative stress.
Service Description
Cell survival rate and intracellular ROS level are the key indicators for screening modeling conditions. The best modeling conditions need to meet that under the treatment concentration and induction time of this stressor, the intracellular ROS level can be significantly increased and the damage to cells can be minimized.
In practical experiments, we usually set up multiple stress source concentrations or intensities and different action time, and comprehensively consider the differences of various indicators under different conditions to select the best modeling conditions. Under certain conditions, the level of intracellular ROS in the model group is significantly higher than that in the normal control group, and the difference is statistically significant. At the same time, the cell survival rate in the model group is not significantly lower than that in the control group. In addition, if there are significant changes in MDA, SOD and GSH PX in the model group, it indicates that the stressor has caused oxidative stress, and the intracellular lipid and enzyme system have changed significantly, the model can be considered successful.
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