Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Rapid primary culture method of haliotis discus hannai gill cells and application

A technology of primary culture and wrinkled abalone, applied in animal cells, invertebrate cells, fermentation, etc., can solve the problems of bacterial pollution and gill cell viability decline, and achieve the effect of solving bacterial pollution and gill cell viability decline

Active Publication Date: 2020-11-03
OCEAN UNIV OF CHINA
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] On the one hand, the present invention provides a rapid primary culture method for gill cells of abalone in wrinkled discs, which can meet the needs of obtaining gill cells of abalone in a short time and then conduct in vitro experiments, solve the problem of bacterial contamination in abalone gill tissue, and solve the problem of using Enzymatic hydrolysis of gill tissue by trypsin enzymatic hydrolysis will cause the decline of gill cell viability

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Rapid primary culture method of haliotis discus hannai gill cells and application
  • Rapid primary culture method of haliotis discus hannai gill cells and application
  • Rapid primary culture method of haliotis discus hannai gill cells and application

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0034] 1.2 Preparation of abalone inactivated serum: about 30 g of blood was collected from the abalone sinus in a wrinkled plate, and it was prepared by filtering and sterilizing through a 0.22 μm membrane filter after bathing in water at 56°C for 30 minutes.

[0035] 1.3 Preparation of complete medium: add 15% fetal bovine serum, 5% inactivated abalone serum, sodium chloride (20.2mg / ml), potassium chloride (0.54mg / ml), chloride Calcium (0.6mg / ml), magnesium sulfate (1mg / ml), magnesium chloride (1.8mg / ml), penicillin (100IU / ml), streptomycin (100μg / ml), gentamicin (250μg / ml), Amphotericin B (2 μg / ml), the dissolved culture medium was sterilized by filtration with a 0.22 μm membrane filter, and placed for later use.

[0036] 1.4 Selection of main reagents: L15 culture medium (Solebol), fetal bovine serum (Biological Industries), penicillin (Solebol), streptomycin (Solebol), gentamicin (Solebol), amphoteric Sodium Chloride (Sigma), Potassium Chloride (Sigma), Calcium Chloride ...

Embodiment 1

[0039] Keep the dissection tools in an autoclave at 121°C for 30 minutes to sterilize and then dry; the dissection tools include tweezers, surgical scissors and scalpels;

[0040] Select the healthy abalone with better vitality as the experimental abalone;

[0041] Clean the body surface of the experimental abalone with a brush, and temporarily raise the experimental abalone with sterile seawater containing disinfectant for 24 hours (the sterile seawater used is prepared in 1.1); Disinfection lamp circulation system for sterilization treatment;

[0042] Wipe the body surface of the experimental abalone clean with 75% alcohol in the aseptic workbench to disinfect the body surface of the experimental abalone; use the above-mentioned sterilized dissection tools to take out the gills of the experimental abalone after disinfection and sterilization, and put into sterile seawater, and wash in a constant temperature shaker at 4°C at a speed of 60rpm for 2h;

[0043] Take the gills ...

Embodiment 2

[0077] This example mainly studies the influence of the centrifugation speed on the experimental results, and whether to select the supernatant or the precipitate after resuspending the complete medium.

[0078] Keep the dissection tools in an autoclave at 121°C for 30 minutes to sterilize and then dry; the dissection tools include tweezers, surgical scissors and scalpels;

[0079] Select the healthy abalone with better vitality as the experimental abalone;

[0080] Clean the body surface of the experimental abalone with a brush, and temporarily raise the experimental abalone with sterile seawater containing disinfectant for 24 hours (the sterile seawater used is prepared in 1.1); Disinfection lamp circulation system for sterilization treatment;

[0081] Wipe the body surface of the experimental abalone clean with 75% alcohol in the aseptic workbench to disinfect the body surface of the experimental abalone; use the above-mentioned sterilized dissection tools to take out the ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a rapid primary culture method of haliotis discus hannai gill cells. The rapid primary culture method comprises the following steps of performing disinfection treatment on experimental abalones; taking out the gills of the disinfected experimental abalones by using a sterilized dissecting tool, and putting the experimental abalones into sterile seawater for cleaning; taking the cleaned gills, cutting the gills into small sections, and adding the gills in a culture dish containing 0.1% of type II collagenase for digestion for 30 minutes; adding the digested gills into aserum-containing culture medium to terminate digestion, performing grinding, performing sieving with a 100 [mu]m cell sieve, and collecting the sieved liquid; centrifuging the sieved liquid, discarding supernatant, performing resuspending with a complete medium, and collecting the supernatant; and inoculating the resuspended and collected supernatant into a T25 culture flask, and performing culturing in an incubator at 22 DEG C;. The method can meet the requirements of obtaining gill cells of the abalones in a short time to perform in vitro experiments, solves the problem of bacterial pollution in gill tissues of the abalones, and solves the problem of gill cell viability reduction caused by enzymolysis of the gill tissues by using a trypsin enzymolysis method. The invention further discloses an application of the method.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a rapid primary culture method and application of gill cells of abalone wrinkled disc. Background technique [0002] Haliotis discus hanai (Haliotis discus hanai) is the main cultured species of marine treasures in my country, and it belongs to the mollusk phylum, gastropod class, abaceae family and genus abalone in taxonomy. Abalone is delicious and nutritious, and is deeply loved by consumers. It is the main abalone cultured in northern my country. At present, the research on wrinkled pan abalone mainly focuses on nutritional requirements and immune stress. However, the breeding cycle of abalone is long, and various parasites are easy to attach to the shell of abalone, which increases the difficulty of carrying out scientific experiments. In order to further study the molecular mechanism of metabolism and immunity of abalone, it is urgent to construct an in vitro model of abalone...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/07C12P21/06
CPCC12N5/0601C12P21/06Y02A40/81
Inventor 张文兵潘明珠刘家欢马硕利刘玥黄冬郭衍林麦康森
Owner OCEAN UNIV OF CHINA
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com