Intestinal Epithelial Cell Modeling Methods
Intestinal epithelial cells are vulnerable to oxidative damage. There are two methods to study intestinal epithelial cell damage: animal model and cell model. Using animal model alone has some disadvantages, such as high cost, instability, difficulty in mechanism research and so on. It is very necessary to construct an appropriate cell culture model as a supplement to the animal model. The methods commonly used by Creative Bioarray to simulate the generation of oxidative stress include physical methods (irradiation, UV irradiation, hypoxia / reoxygenation) and chemical methods (hydrogen peroxide modeling, xanthine / xanthine oxidase modeling).
The following are the oxidative stressors we can provide at present. Each modeling method has its applicable simulation conditions. We will implement the most appropriate modeling scheme according to the actual research content of customers.
Establishment of Epithelial Cell Oxidative Stress Model by Physical Methods
Irradiation
Oxidative stress is mediated mainly by reactive oxygen species produced by ionization. After irradiation, the intestinal tract of animals was collected for oxygen free radical and cell damage analysis. However, compared with cell experiments, this method is more suitable for animal level experiments. Therefore, we suggest that customers can choose this method as a model for studying the radiation damage of antioxidants to intestinal epithelium.
Hypoxia / reoxygenation
The basic principle of hypoxia / reoxygenation operation is to simulate the process of ischemia-reperfusion.
The usual method is to place the cultured cells in anaerobic environment for several hours, and then return to the culture state of normal oxygen supply for a period of time, so as to cause oxidative damage to the cells.
We suck the subcultured cells out of the culture medium, place them in an anaerobic environment (e.g. 1% O2, 5% CO2, 94% N2), and take them out after incubation for an appropriate time. After adding the medium, it was placed in an aerobic environment (5% CO2, 95% O2) to cause oxidative damage.
Chemical Methods—H2O2 Modeling Service
H2O2 is relatively stable compared with other ROS. It can construct a severe and stable cellular oxidative stress model in intestinal epithelial cells. Hydrogen peroxide can generate high activity-OH (H2O2 + Fe2+ + H+ → - OH + Fe3+ + H2O) through Fenton reaction in the presence of ferrous Homo environment in vivo.
When modeling, we usually consider the amount of H2O2 and treatment time from three aspects.
First simulated object
The template treated with H2O2 can simulate acute oxidative damage within 3h. The model with treatment time greater than 12h can be used to study the long-term stress process.
Cell type
Different cell types have different sensitivity to H2O2. Primary cultured intestinal epithelial cells are usually modeled with a smaller dose, while higher doses of H2O2 are required for subculture cell modeling.
Actual situation of experiment
In short-term modeling, the dose is too small to have significant difference. If the dose is too large, a large number of cells will die and float in the culture medium, causing trouble for the follow-up test. We will explore according to the customer's research object and test conditions to explore the best modeling dose and time.
Creative Bioarray is dedicated to providing high-quality products, comprehensive services, and tailored solutions to support and facilitate life sciences and pharmaceutical research and development. If you have any questions or needs, please contact us, and our customer service staff will help you at the first time.
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