Digestive juice for cartilage tissues, and use method thereof

Abstract

The invention discloses a digestive juice for cartilage tissues. The digestive juice comprises the following components: 1-3 mg / ml of type II collagenase; 1-3 mg / ml of type IV collagenase; 5 ml of neutral protease; 12 units / ml of DNA enzyme; 10% of FBS (V / V); and 0.053 mg / L of CaCl2. The beneficial effects are as follows: a cell acquisition rate is high, cells have relatively high activity, a clustering rate is low, the damage to the cells is relatively small, the transcriptome change is low, and the subsequent culture or single-cell transcriptome sequencing can be performed.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a digestive juice for cartilage tissue and a digestion method thereof. Background technique [0002] Cartilage tissue is composed of chondrocytes, fibers and matrix. During the period of embryogenesis, cartilage acts as a temporary bone and becomes the frame of the body. With the development of the fetus, the cartilage is gradually replaced by bone, and some cartilage tissue is still retained in the adult body, which has the function of support and protection. [0003] In recent years, many studies have shown that chondrocyte dedifferentiation is an important phenomenon in the pathology and repair of articular cartilage damage. Many research methods require cartilage tissue to be digested into single cells for research, such as cartilage tissue engineering research, chondrocyte differentiation research, etc. Therefore, cartilage The method of culturing cells becomes one of...

AI Technical Summary

Problems solved by technology

[0006] 1. Although type II collagenase is a mild digestive enzyme, it still damages cells, especially when it is over-digested, the damage to cells is very obvious

[0007] 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 chondrocytes

[0008] 3. Chondrocytes are prone to incomplete digestion after digestion with type II collagenase, and it is easy to form 2 cell clusters. For some follow-up experiments such as single-cell transcriptome sequencing, clustered cells will lead to failure of subsequent experiments

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