Cardiomyocyte model of heart qi deficiency in mice
Cardiomyocyte model of heart qi deficiency in mice
The cardiomyocyte model of heart qi deficiency mice is based on hypoxia as a simulated etiology of heart qi deficiency evidence, which leads to ischemia of the myocardium and impairment of cardiac function. Currently, the cardiomyocyte model of heart qi deficiency mice is established by the method of hypoxia and reoxygenation injury of cardiomyocytes.
Principle
The modeling principle of the cardiomyocyte model of cardiac qi deficiency mice is ischemia of the myocardium, hypoxia can often lead to cardiac function damage, hypoxia as the simulated etiology of cardiac qi deficiency, the use of cardiomyocyte hypoxia and then oxygen damage to the establishment of the cardiac qi deficiency evidence cell model.
Appliance
The cardiomyocyte model of heart qi deficiency mice can be combined with the overall animal model of heart qi deficiency evidence to directly reflect the pathological state in vivo, reflecting the characteristics of Chinese medicine.
Operation method
Establishment of a cardiomyocyte model in cardiac qi deficient mice by hypoxia-reoxygenation injury method in cardiomyocytes
Principle
The basic principle of the cardiomyocyte hypoxia-reoxygenation injury method to establish a cardiomyocyte model of cardiac qi deficiency in mice is to use hypoxia as a simulated etiology of cardiac qi deficiency evidence, which leads to ischemia of the myocardium and impairment of cardiac function.
Materials and Instruments
Experimental animals: NIH mouse suckling mice. Move The modeling method for establishing cardiomyocyte model of cardiac qi deficiency mice by cardiomyocyte hypoxia and reoxygenation injury method is as follows: A. The skin was sterilized with 75% ethanol, the chest cavity was cut open with iris clippers, and the heart of the suckling mouse was clamped with tissue forceps, placed in Hanks' solution for washing, the atria were clipped, and then rinsed repeatedly with Hanks' solution to wash the residual accumulated blood. B. Place the ventricles of suckling rats on the side wall of the centrifuge tube, cut each ventricle with iris scissors, add 0.1% trypsin 10 ml, and vibrate the digested tissue for 10 minutes in a 37 ℃ water bath, and then discard the supernatant after natural precipitation, then add trypsin 10 ml, and then vibrate the digested tissue again for 10 minutes in a 37 ℃ water bath. After the tissue was blown by a pipette for 1 minute, it was allowed to settle naturally. C. Transfer the supernatant to another sterile centrifuge tube and add 2 ml of culture medium to terminate digestion. The precipitate was continued to be digested by adding trypsin as before, and the supernatant was collected as before, and this was repeated 6 times. D, the supernatant was centrifuged continuously at 5000 r/min for 5 min, and the supernatant was discarded, and then the centrifugation was repeated for 1 time with culture medium. The cell suspension was placed in a culture flask and allowed to stand for 1.5 hours at 37 ℃ in a constant temperature incubator, and then the cardiomyocyte suspension was aspirated and inoculated into a 24-well culture plate of Costar after counting with a leukocyte counter. E. 2. 5×105 cardiomyocytes were inoculated in each well, and 1 ml of 1640 medium containing 20% calf serum was added to each well, and the plate was sealed with transparent adhesive and placed in 37 ℃ constant temperature incubator for culture, and the medium was changed once after 2 days. F. Take a monolayer of cardiomyocytes after 3 days of culture for the experiment, place the culture plate in a container, and use 99.99% high-purity nitrogen to inflate the container for 1 minute and then close the mouth of the container tightly. For more product details, please visit Aladdin Scientific website.
Reagents: Hanks solution.