High-purity cardiac muscle cell primary culture method

Abstract

The invention provides a high-purity cardiac muscle cell primary culture method and is suitable for the field of biotechnology and medical science. The high-purity cardiac muscle cell primary culture method particularly comprises the steps that an isolated heart of a suckling mouse is sheared into tissue fragments; digestion is conducted on the tissue fragments through improved prepared enzyme digestive juice for multiple times, cardiac muscle cell suspension is prepared, resuspension is conducted, and then cardiac muscle cells are inoculated in a culture dish A for differential-time wall sticking; cardiac muscle cell suspension in the culture dish A is sucked into a culture dish B, cardiac muscle cells which sink to the bottom of the culture dish A and do not stick to the wall in the culture dish A are cleaned and resuspended, and then the resuspended cardiac muscle cells are transferred into the culture dish B to be cultured. Compared with the prior art, the high-purity cardiac muscle cell primary culture method has the advantages that the method is simple and effective, the operability is strong, the number of obtained cardiac muscle cells is large, the purity is high, the wall sticking rate is good, and the survival rate is high.

Description

technical field [0001] The invention provides a primary culture method of high-purity cardiomyocytes, which is suitable for the research on related cardiovascular diseases at the cell level, and clarifies the mechanism problem from the cell molecular level, and belongs to the fields of biotechnology and medicine. Background technique [0002] As the incidence of heart disease increases year by year, the research on myocardial disease is getting more and more attention. The in vitro culture of neonatal rat cardiomyocytes can retain some of the original structural and functional characteristics of the body, has pulsation, and is not disturbed by factors such as nerves and body fluids, and can be used for cardiomyocyte growth and development, pathophysiology, and pharmacology. Therefore, the primary culture of neonatal rat cardiomyocytes is widely used in the field of prevention and treatment of cardiovascular diseases. [0003] However, due to the various primary culture meth...

AI Technical Summary

Problems solved by technology

[0007] Disadvantages: The number of cardiomyocytes obtained is small, which is difficult to meet the needs of the experiment

[0010] Disadvantages: greater damage to cells, due to the action of enzymes on cardiomyocytes, as well as centrifugation, resuspension and other steps, there will be different degrees of chemical or physical damage to isolated cardiomyocytes

Fibroblasts in heart tissue have strong wall-attachment ability, and cardiomyocytes are damaged to varying degrees under the digestion of enzyme solution, so it is easy to form the growth advantage of fibroblasts, thereby gradually diluting cardiomyocytes, resulting in the isolated The purity of cardiomyocytes is very low

[0011] Both methods have their own advantages and disadvantages. Therefore, it is a technical problem to obtain a large number of cardiomyocytes with high survival rate and high purity in a short period of time to meet the experimental requirements.

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