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A Gift Tilapia Brain Neural Cell Line, Transfection Method, Culture Method and Application

A technology of tilapia and brain nerve cells of GIFT, applied in the biological field, can solve the problems of long time and the inability to obtain commercial cell lines, and achieve the effect of easy transfection

Active Publication Date: 2022-02-22
INST OF AQUATIC LIFE ACAD SINICA
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The main purpose of the researchers to establish these tilapia cell lines is to isolate and cultivate viruses, and these cell lines are not commercially available, and some cell lines have been reported for a long time

Method used

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  • A Gift Tilapia Brain Neural Cell Line, Transfection Method, Culture Method and Application
  • A Gift Tilapia Brain Neural Cell Line, Transfection Method, Culture Method and Application
  • A Gift Tilapia Brain Neural Cell Line, Transfection Method, Culture Method and Application

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Experimental program
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Embodiment

[0024] 1. Breeding of Experimental Fish

[0025] The fry of Gifu tilapia were purchased from the fish farm of the Freshwater Fisheries Research Center of the Chinese Academy of Fishery Sciences. After being transported back to the laboratory, they were cultured in the indoor circulating water system, the water temperature was controlled at 28±0.5°C, and the lights were illuminated for 12 hours (8:00-20:00) every day. According to the size, they were fed Artemia, frozen red worms and floating pellet feeds in turn.

[0026] 2. Primary Cell Culture

[0027] (1) A juvenile Gifu tilapia with a body length of 5.2 cm and a body weight of 5.72 g was anesthetized with 80 mg / L MS-222 (Sigma-Aldrich) and dissected.

[0028] The following operations are all carried out in a biological safety cabinet, using sterile consumables and reagents dedicated to cell culture.

[0029] (2) Take out the brain tissue and soak it in 70% alcohol for 5 sec.

[0030] (3) Rinse the tissue pieces in 5 mL...

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Abstract

The invention belongs to the field of biotechnology, and in particular relates to a brain nerve cell line of a gift from tilapia, a transfection method, a culture method and applications thereof. In the present application, an immortal cell line was isolated and cultured from the brain tissue of Gift Tilapia, named TBN. The cell line has neuron-like morphological characteristics, expresses a series of neuronal marker genes, and is a neural precursor cell line; it is sensitive to low temperature and cannot survive under low temperature conditions of 16 °C and below; and it is easy to transfect , the transfection efficiency of electroporation method is as high as 52%. This cell line can be used as a good model to study the function of cold resistance-related genes in tilapia, filling the gap in related fields.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a gifu tilapia brain nerve cell line, a transfection method, a culture method and an application thereof. Background technique [0002] Tilapia (Oreochromis sp.) has excellent characteristics such as fast growth, strong disease resistance, and rich nutrition, and is one of the important aquaculture species. However, its low temperature tolerance has led to severe restrictions on the breeding area, increased breeding costs, and reduced breeding benefits. Due to the large size of tilapia and its habit of building nests and hatching fertilized eggs in its mouth, it is not easy to perform operations such as microinjection, and it is difficult to carry out targeted gene function research as efficiently as research models such as zebrafish (Danio rerio) , so the current understanding of the genetic determination basis of cold tolerance traits in tilapia is still insufficient. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/079C12N15/87C12Q1/02C12R1/91
CPCC12N5/0618C12N15/87G01N33/5005C12N2500/84
Inventor 龙勇刘冉宋桂丽李青崔宗斌
Owner INST OF AQUATIC LIFE ACAD SINICA
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