Construction method of mandarin fish brain cell line

Abstract

The invention discloses a construction method of a mandarin fish brain cell system. Aiming at the characteristic of mandarin fish brain cells, the primary cell culture is carried out by separating the brain cells through a serum-contained collagenase digestion method, so that the mandarin fish brain cell system is constructed successfully. Thus, a great quantity of mandarin fish brain source cells can be provided; and the good growth status of sub-culture cells can be maintained. The construction method is strong in repeatability and brings convenience for researching fish virus infection paths and fish virus infection mechanisms, developing viral vaccines and the like.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular to a method for constructing a mandarin fish brain cell line. Background technique [0002] Mandarin mandarin fish (Siniperca chuatsi), also known as osmanthus fish, has tender meat and delicious taste. It is known as "freshwater grouper" in the international aquatic product market. It is one of the important varieties of high-quality freshwater fish consumption and export earnings in my country. With the expansion of mandarin fish breeding scale and the increase of breeding density, there are many diseases of mandarin fish. The pathogens include viruses, bacteria, parasites, etc., especially the viral diseases caused by infectious spleen and kidney necrosis virus (ISKNV) are the most serious. The rate is generally more than 60%, and the mortality rate is more than 50%. When it is serious, the mortality rate of mandarin fish in ponds reaches 90%, or even all of them die. sustainable de...

AI Technical Summary

Problems solved by technology

However, many brain tissue cells of mature fish have not divided vigorously or have not divided, and in the current fish cell line construction method, the rapid shearing tissue method will generate mechanical shear force, which will have a great impact on brain cells, making It is difficult for primary brain cells to attach and proliferate normally, and the trypsin digestion separation method will cause a large number of brain cells to die due to hypoxia during trypsin digestion. Therefore, it is necessary to study an effective method for the construction of mandarin fish brain cell lines and establish The mandarin fish brain cell line lays the foundation for studying the pathogenic mechanism of viruses and developing antiviral drugs and antiviral vaccines at the cellular level

So far, there have been reports in the literature that a juvenile mandarin fish cell line was established by using trypsin digestion and isolation (Sun Yat-Sen University, Dong Chuanfu, Virus Research, 2008, 135: 273–281), but there is no report on a brain-derived cell line

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