Haliotis discus hannai cell culture medium and construction method of cell line

A technology of wrinkled plate abalone and construction method, which is applied in the direction of cell culture active agent, microorganism-based method, culture process, etc., can solve the problems of affecting cell immunology, poor cell morphology, short survival time, etc., and is beneficial to popularization and The effect of popularization, long cultivation time and good growth state

Active Publication Date: 2020-01-14
福建罗屿岛食品有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In view of many problems such as poor cell morphology, low survival rate, short survival time, and difficulty in passage in the culture of Abalone wrinkled cells, it has seriously affected the research on cellular immunology, virology, develop

Method used

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  • Haliotis discus hannai cell culture medium and construction method of cell line
  • Haliotis discus hannai cell culture medium and construction method of cell line
  • Haliotis discus hannai cell culture medium and construction method of cell line

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Embodiment 1

[0033] Prepare abalone cell culture medium according to the following formula: use L-15 medium as the base medium, and add the following components at the final concentration: NaCl 12.0g / L, KCl 0.45g / L, ZnSO 4 0.3g / L, MgCl 2 1.0g / L, CaCl 2 0.15g / L, KH 2 PO 4 60mg / L, glucose 1.0g / L, basic fibroblast growth factor 17.5ng / mL, epidermal growth factor 4ng / mL, L-glutamine 0.3g / L, astaxanthin 25μmol / L, γ-aminobutyric acid 18mg / L, carboxymethyl dextran 50mg / L, taurine 40mmol / L, hydrocortisone 2.0mg / L, abalone shell water extract 60mg / L, fetal bovine serum 10%, penicillin sodium 100IU / mL, streptomycin sulfate 100 μg / mL, imipenem 10 μg / mL. The pH value of the medium was adjusted to 7.2, and the osmotic pressure was adjusted to 1000 mOsm / kg. After the preparation was completed, it was sterilized by filtration with a 0.22 μm filter membrane, and refrigerated at 4°C for later use.

[0034] The preparation method of the water extract of the wrinkled disc abalone shell is as follows...

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Abstract

The invention provides a haliotis discus hannai cell culture medium, which is based on an L-15 culture medium and is added with inorganic salt, growth factors, L-glutamine, astaxanthin, gamma-aminobutyric acid, carboxymethyl glucan, taurine, hydrocortisone, a haliotis discus hannai shell aqueous extract, fetal calf serum, antibiotics and other components. The invention also provides a constructionmethod of the haliotis discus hannai cell line, which comprises the following steps: by using haliotis discus hannai heart tissues as a material, obtaining single cells by a mechanical dissociation method, and resuspending the single cells in the cell culture medium for primary culture, thereby successfully constructing the haliotis discus hannai heart cell line. The culture medium and the methodprovided by the invention can well maintain survival and growth of haliotis discus hannai heart cells in vitro, and the constructed cell line has continuous passage capacity. According to the method,the survival rate of the haliotis discus hannai heart cells in-vitro culture is remarkably increased, the in-vitro culture time is prolonged, and the method has the characteristics of quickness, easiness in operation, stability and easiness in repetition.

Description

technical field [0001] The invention belongs to the technical field of animal cell culture, and in particular relates to a cell culture medium and a method for constructing a cell line of abalone wrinkled. Background technique [0002] Abalone (Abalone), also known as sea ear, bream, and mirror fish, belongs to the mollusk phylum, gastropods, primitive gastropods, abaceae, and genus Abalone. It is an algae-eating shellfish and usually grows on the seabed with low water temperature. Footprints throughout the Pacific, Atlantic and Indian Oceans. Abalone is a narrow-temperature and narrow-saline shellfish. The environment in which it lives in the sea area requires clear water quality and smooth currents, and the salinity of seawater is kept above 3% all year round. Abalone has been hailed as the crown of seafood treasures since ancient times. It is known as "one bite of abalone, one bite of gold". Its meat is tender, delicious, and has extremely high nutritional value. Abalon...

Claims

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Application Information

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IPC IPC(8): C12N5/07C12R1/91
CPCC12N5/0601C12N2500/12C12N2500/14C12N2500/16C12N2500/22C12N2500/30C12N2500/32C12N2500/33C12N2500/34C12N2500/82C12N2501/11C12N2501/115C12N2501/39
Inventor 唐金锁胡晓明庄涛
Owner 福建罗屿岛食品有限公司
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