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A method for the isolation and primary culture of Chinese mitten crab hepatopancreatic cells

A Chinese mitten crab, primary culture technology, applied in the field of cell culture

Active Publication Date: 2020-04-28
XICHANG COLLEGE
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, in the prior art, there are no reports about the separation and primary culture method of the Chinese mitten crab hepatopancreas of the present invention.

Method used

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  • A method for the isolation and primary culture of Chinese mitten crab hepatopancreatic cells
  • A method for the isolation and primary culture of Chinese mitten crab hepatopancreatic cells
  • A method for the isolation and primary culture of Chinese mitten crab hepatopancreatic cells

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Embodiment 1

[0025] 1 Materials and methods

[0026] 1.1 Test material

[0027] Chinese mitten crabs are collected from the Aquatic Technology Promotion Station of Jintan City, Jiangsu Province. The size is 125g~135g / piece, male or female, and they are temporarily kept in a 75cm×50cm×45cm breeding box. The photoperiod is 12h: 12h, and the water is changed every day. 3. Feed the compound feed at a fixed time in the morning and evening, and suck out the residual bait and feces in the tank every day to ensure the water quality is clear.

[0028] Crab saline (crab saline): NaCl (14.5g), KCl (0.355g), anhydrous CaCl 2 (0.899g), MgSO 4 ·7H 2 O (1.58g), NaHCO 3 (0.25g), MgCl 2 ·6H 2 O (0.85g), HEPES (2.38g), distilled water (500ml).

[0029] 2×L-15 and 3×L-15 culture medium: 1L specification L-15 dry powder (Gibico) was dissolved in 500ml and 333ml distilled water respectively, sterilized by 0.22μm filter membrane in a sterile environment, and then dispensed into sterile In a blue cap bot...

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Abstract

The invention relates to an Eriocheir hepatopancreas cell separation and primary culture method. The method comprises the following steps: (1) cleaning Eriocheir pancreatic tissues with cell separation liquid, cutting the cleaned Eriocheir pancreatic tissues to tissue blocks of 1mm<2> with a shear, adding a proper quantity of tissue blocks into the separation liquid, blowing and filtering with a suction tube, and centrifuging the filtrate to obtain yellow precipitate; (2) adding the yellow precipitate into a culture medium containing 100UI / ml penicillin and 100mug / ml streptomycin, blowing uniformly, inoculating into a culture plate, and culturing in a CO2 cell incubator. By adopting the method, Eriocheir hepatopancreas cells can be stably cultured till the survival rate with 72 hours is higher than 50 percent, a theoretical basis is laid for building of a subsequent cell line, and a cell tool is provided for researches in relevant fields of cell physiology, gene engineering and the like.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to a method for the separation and primary culture of Chinese mitten crab hepatopancreatic cells. Background technique [0002] The Chinese mitten crab (Eriocheir) belongs to the decapod crustacea and is one of the main aquaculture species in my country's aquaculture industry. With the expansion of breeding scale and the increase of breeding years, the outbreak of various diseases, especially some viral diseases, has had a huge impact on the Chinese mitten crab farming industry. The hepatopancreas is an important tissue and organ of crabs. It is responsible for many physiological functions, including synthesizing and secreting digestive enzymes, absorbing nutrients, storing inorganic substances, and metabolizing carbohydrates. In addition, the hepatopancreas is also involved in excretion and molting. Activity. The hepatopancreas is composed of a variety of cells, which can be...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071
CPCC12N5/0676C12N2509/00
Inventor 洪宇航杨筱珍成永旭黄毅
Owner XICHANG COLLEGE
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