Method for preparing high-quality single-cell suspension by remarkably improving plaque digestion

Abstract

The invention discloses an artery plaque digestion method capable of remarkably improving the quality of a single-cell suspension. The kit comprises a collagen hydrolase I, a collagen hydrolase II, acollagen hydrolase IV, a collagen hydrolase XI, calcium chloride, deoxyribonuclease I, a phosphate buffer salt solution, a bovine serum albumin solution and an RPMI 1640 cell culture solution. The invention also discloses an application prospect of the polypeptide in digesting plaque tissues. The invention is invented mainly for digestion treatment of plaque tissues. Through combination of severalenzymes, plaque tissue can be digested into a single-cell suspension with a relatively high activity proportion, and the biological reaction of the plaque tissue is researched on the basis of singlecells, so that the situation that a large number of calcified cell debris, tissue impurities and the like influence a single-cell sequencing machine or a subsequent data analysis result is avoided.

Description

technical field [0001] The invention relates to the field of plaque digestion methods, in particular to the plaque tissue digestion method to prepare high-quality single cell suspension and its application. Background technique [0002] The "China Cardiovascular Disease Report 2018" shows that the prevalence and mortality of cardiovascular and cerebrovascular diseases in China are still on the rise. At present, cardiovascular and cerebrovascular diseases have become a major public health problem, and the control of atherosclerosis is closely related to the prevention and treatment of cardiovascular and cerebrovascular diseases. Atherosclerosis is the main pathological basis of cardiovascular and cerebrovascular diseases. Risk factors such as high blood pressure, smoking, and hyperhomocysteinemia can all cause atherosclerosis in the body, and even due to maternal hypercholesterolemia, the earliest The primary state of atherosclerosis, "lipid streaks", can be observed in the ...

AI Technical Summary

Problems solved by technology

[0003] In the process of plaque processing, if mechanical mincing and enzymatic digestion are not handled well, many cell debris and a large number of dead cells will appear, resulting in long operation time when filtering cell suspension and subsequent single-cell transcriptome sequencing When the machine is on, the pipeline will be blocked, which will cause damage to the precision instrument.

Although there are many tissue digestion kits on the market, due to the particularity of the plaque, the digestion is not complete, there are a lot of cell debris and dead cells, the digestion effect cannot be achieved, and the quality of the prepared single-cell suspension cannot be obtained. ensure

At present, the problem of thorough digestion of plaque tissue and elimination of calcified debris and cell debris has not been well resolved

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