Construction method of muscle cell line of anabarilius grahami

Abstract

The invention relates to a construction method of a muscle cell line of anabarilius grahami and belongs to the technical field of freshwater organism cell culture. The method comprises the steps of firstly preparing cell culture fluid, ensuring that anabarilius grahami muscle tissue serves as a material, inoculating in a culture bottle of 25 square centimeters, and placing a culture box at a temperature of 28 DEG C for culture; replacing the culture liquid in half volume every 3 days, and performing passage by using a trypsin digestion method after a cell grows into a monolayer; and performing passage every 5 days, when the passage of a tenth generation is achieved, the serum concentration is the culture fluid is reduced to 10%, and the cell line is constructed successfully. The method has the advantages that 1, the operation is easy and feasible; 2, in the constructed anabarilius grahami muscle cell line (AGM), the cell growth is good, the cells are in a fibroblast cell pattern, above 50 generations can be passed to continuously, and the method can be applied to studies of anabarilius grahami cell biological characteristics and immune resistance gene functions directly; and 3, the method is also applicable to muscle cell lines of other fishes.

Description

Technical field:

[0001] The invention relates to a method for constructing a muscle cell line of the whitefish, belonging to freshwater aquatic organism cell culture technology. Background technique:

[0002] Fish cell culture began in the early 1960s, when Clem et al. began to study the tissue culture of seawater fish. Up to now, there have been more than 250 cell lines. The culture of fish cells in my country started in the 1970s. Most of the early research focused on freshwater cultured fish, especially carps. So far, more than 40 cell lines have been established, mainly from more than 20 kinds of fish. However, there are very few indigenous fishes in Yunnan.

[0003] Anabarilius grahami belongs to the order Cypriniforms, the family Cyprinidae, and the genus Anabarilius, commonly known as white fish. It is a rare fish species only produced in Fuxian Lake, Yunnan, and it is one of the four famous fish in Yunnan. The fish is tender and delicious, with soft spines and thin ...