Isolation and culture method for primary mice or rat cartilage cells

Abstract

The invention discloses an isolation and culture method for primary mice or rat cartilage cells. According to the method, a step of polishing the surface of cartilage is carried out in acquisition of the cartilage cells, so contamination by subsequent synovial cells is maximally eliminated; the method innovatively employs a biopsy needle to cut interior cartilage tissue, so sampling is directed at target cells, the method is direct and effective, and potential bacterial and fungal contamination is ingeniously prevented; meanwhile, the isolation and culture method employs a specially-prepared compound enzyme digestive juice, so compared with traditional single collagenase preheating and digestion, the method provided by the invention overcomes the problems of low cell viability and poor yield caused by excess digestion and excess blowing and beating of tissue, and enables digestion to be more uniform and thorough.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for isolating and culturing primary mouse or rat chondrocytes. Background technique [0002] Chondrocytes are located in cartilage, and their main function is to synthesize some collagen and non-collagenous macromolecular extracellular matrix, including: type II collagen, proteoglycan, link protein, type IX collagen and type XI collagen. Regulating the proliferation and differentiation of chondrocytes is critical for coordinating skeletal development in vertebrates. Chondrocytes can produce and react with a large number of peptide growth factors and cytokines, such as insulin-like growth factor-1 and interleukin-1. The culture of chondrocytes is a very important in vitro model in the study of cartilage regeneration and repair, the effects of cytokines and growth factors on cartilage, the regulation of specific genes on arthritis, and the pathophysiological process ...

AI Technical Summary

Problems solved by technology

The disadvantage is that most of the enzymes will undergo severe digestion at 37 degrees. Therefore, on the one hand, it can speed up the speed of digestion, but on the other hand, it will cause great damage to the cells, especially the pancreas. Enzyme even more so

As a result, the digestion time is not easy to grasp, often the digestion time is too long, the cell fragments are mostly, there are more miscellaneous cells and less chondrocytes, and the obtained cells have poor viability. There are fewer and fewer chondrocytes, and the cells gradually die

At the same time, during the collection of chondrocytes, the contamination of other cells such as synoviocytes is very common. In addition to the above-mentioned enzyme digestion process, it is easy to cause the number of chondrocytes to be less dominant, while other miscellaneous cells with higher tolerance are isolated instead. more

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