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Construction method of mandarin fish brain cell system

A construction method and technology of brain cells, applied in artificial cell constructs, methods based on microorganisms, animal cells, etc., can solve the problem of brain cell death due to hypoxia, no reports of brain-derived cell lines, and the difficulty of normal adhesion of primary brain cells wall proliferation

Active Publication Date: 2013-09-04
PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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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

Method used

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  • Construction method of mandarin fish brain cell system
  • Construction method of mandarin fish brain cell system
  • Construction method of mandarin fish brain cell system

Examples

Experimental program
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Embodiment 1

[0040] 1) Treatment of brain tissue: First, soak and disinfect fresh mandarin fish with 75% alcohol for 2 minutes, wipe off the mucus on the body surface with gauze, dissect and remove the brain tissue in an ultra-clean workbench, and put it in a small chamber filled with HBSS balance solution. The brain tissue was roughly cut into tissue pieces of 50-100 mm3 in a glass bottle, collected by centrifugation at 600 rpm, and centrifuged repeatedly for 3 times.

[0041] 2) Primary culture: put the tissue block into the serum-containing collagenase digestion solution (digestion solution contains 10v / v% fetal bovine serum, 0.1w / v% type Ⅰ collagenase, 200U / ml penicillin, 200μg / ml ml streptomycin, 0.5 μg / ml amphotericin B in L-15 medium) for 6 hours at 28°C, after digestion, centrifuge at 1200 rpm for 5 minutes, collect the digested tissue cell suspension, Add 5 ml special proliferation culture medium for mandarin fish brain cells (the formula of culture medium is containing 20v / v% fet...

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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...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071C12R1/91
Inventor 付小哲李宁求吴淑勤罗霞赖迎迢石存斌
Owner PEARL RIVER FISHERY RES INST CHINESE ACAD OF FISHERY SCI
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