Digestive enzyme liquid for poultry tibial growth plate cartilage primary cell separation and separation culture method of tibial growth plate cartilage primary cells

Abstract

The invention provides digestive enzyme liquid for poultry tibial growth plate cartilage primary cell separation. A complete medium is used as basic liquid, and is prepared from the following ingredients including collagenase IV with the mass concentration being 80 to 100mg/mL and hyaluronidase with the mass concentration being 80 to 100mg/mL; the complete medium takes a high-glucose DMEM-F12 culture medium as a basic culture medium, and contains the following ingredients including fetal calf serum with the volume concentration being 8 to 12 percent, vitamin C with the mass concentration being 4.5 to 5.5mg/mL and mycillin with the mass concentration being 1 to 10mg/mL. Through the selection on digestive enzyme and the concentration optimization, the cell yield reaches 85 to 95 percent or higher; during the further culture on the digested cells, the cell upward trend is good; meanwhile, the cell viability of the digested primary cell reaches 85 to 95 percent or higher; the cell viability is relatively stable.

Description

technical field [0001] The invention belongs to the technical field of cell culture, and in particular relates to a digestive enzyme solution for isolating poultry tibia growth plate cartilage primary cells and a method for separating and culturing the tibial growth plate cartilage primary cells. Background technique [0002] Tibial chondrodysplasia (TD) is a cartilage metabolic disease that occurs in poultry. The disease often occurs in poultry such as broiler hens, layers and turkeys. The clinical manifestations are depression, loss of appetite, often crawling on the bottom, dyskinesia, difficulty standing, and sometimes walking with the support of wings. TD can reduce the disease resistance and production performance of chickens, induce breast cysts and osteomyelitis, reduce the quality and grade of chicken meat, endanger animal welfare, and cause important losses to the field of poultry farming. It occurs in all countries in the world, and causes huge economic losses to...

AI Technical Summary

Problems solved by technology

However, the cause of the decline in viability and death in the in vitro culture process is still unclear. Therefore, a complete set of isolation and culture methods have not been successfully formed in the prior art.

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