Tumor tissue digestive juice and method thereof

Abstract

The invention discloses tumor tissue digestive juice and a method thereof. The digestive juice comprises the following components: type II collagenase of 2 mg / ml, type IV collagenase of 2 mg / ml, Dispase (neutral protease) of 0.2 ml / ml, DNA enzyme of 12units / ml, FBS of which the volume fraction of is 10%, DMEM of 0.8ml / ml, and CaCl2 of 3mM. According to the tumor tissue digestive juice provided bythe invention, the Dispase (neutral protease) is added, the protease can better digest and dissociate protein connection between compact tumor cells, besides, the Dispase enzyme content is increased,and the digestion efficiency is remarkably improved. Besides, the type IV collagenase is added to the tumor tissue digestive juice provided by the invention, so that the digestive efficiency can be remarkably improved, the digestive time can be shortened, the concentrations of the type IV collagenase and the type II collagenase in the digestive juice are relatively low, the cell damage can be reduced, and the cell viability in a cell suspension can be improved.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a tumor tissue digestive fluid and a method thereof. Background technique [0002] With the development of biomedicine, in order to solve the problem that gene high-throughput sequencing technology conceals the heterogeneity between cells in the process of oncology research, since single-cell transcriptome sequencing technology can reveal tumor tissues at the single-cell level The characteristics and interconnections of different cells, in order to study the biological characteristics of a single tumor cell, the single-cell transcriptome sequencing technology of tumor cells has become the focus of clinical experts and scientific researchers in oncology. [0003] Single-cell suspensions are widely used in the research of tumor pathology and tumor molecular biology. At present, the commonly used preparation methods for single-cell suspensions of fresh tumor tissues are: firstly...

AI Technical Summary

Problems solved by technology

The enzyme schemes currently used for digestion of different tumor tissues may be different. Sensitive cell types may be destroyed during different cell digestion processes. Some tumor tissues have low tissue content after surgery, resulting in insufficient digestion of the cell suspension after digestion, which affects follow-up experiments

[0004] At present, in the process of tumor digestion, the existing problems mainly include: incomplete tissue digestion, long digestion time, high enzyme concentration, etc., which lead to cell death, resulting in a series of problems in subsequent experiments

[0005] At present, the tumor cell separation method will be commonly obtained by type II collagenase digestion method, but this method has many shortcomings, mainly including: 1. Although type II collagenase is a mild digestive enzyme, it still has a strong effect on cells. Damaged, especially when overdigested, the damage to cells is evident

2. Type II collagenase is prone to loss of activity during use, which will reduce the digestion effect, and in serious cases will lead to the inability to realize the digestion and separation of tumor cells

3. Tumor cells are prone to incomplete digestion after digestion with type II collagenase, and the digestion time is too long, which will lead to a more serious impact on subsequent single-cell sequencing experiments

Images