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Method for culturing grouper cells by using bioreactor

A bioreactor, grouper technology, applied in the biological field, achieves the effect of high cell yield, normal cell shape, and promotion of low biological metabolism

Active Publication Date: 2021-09-28
GUANGZHOU QIZHI BIOLOGICAL ENG EQUIP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are few methods of using bioreactors to cultivate grouper cells at home and abroad

Method used

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  • Method for culturing grouper cells by using bioreactor
  • Method for culturing grouper cells by using bioreactor
  • Method for culturing grouper cells by using bioreactor

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Embodiment 1 uses BC-7L bioreactor suspension to cultivate grouper cell method

[0032] The method for cultivating grouper cells using BC-7L bioreactor comprises the following steps:

[0033] 1) The grouper GS cells were digested with 0.25% trypsin digestion solution to prepare a cell suspension; the trypsin digestion solution included 0.02% ethylenediaminetetraacetic acid and 0.25% trypsin (1 :250);

[0034] 2) Inoculate the cell suspension prepared in step 1) into a BC-7L bioreactor containing cell growth fluid and microcarriers for dynamic culture; the amount of cell growth fluid added is 1000ml; including 800ml basal medium, 50ml Galactose, 150ml grouper tissue fluid; the basal medium is DMEM basal medium, and the preparation process of the grouper tissue fluid is: add 100ml normal saline to 10g grouper tissue, grind and stir, and centrifuge the supernatant at 10000rpm Liquid, sterilized and filtered, to get it; the microcarrier is Cytodex1; the addition amount is...

Embodiment 2

[0036] Embodiment 2 uses BC-7L bioreactor suspension culture grouper cell method

[0037] The method for cultivating grouper cells using BC-7L bioreactor comprises the following steps:

[0038] 1) The grouper GS cells were digested with 0.25% trypsin digestion solution to prepare a cell suspension; the trypsin digestion solution, in terms of mass fraction, included 0.02% ethylenediaminetetraacetic acid and 0.25% trypsin ( 1:250);

[0039] 2) Inoculate the cell suspension prepared in step 1) into the BC-7L bioreactor containing cell growth liquid and microcarriers for dynamic culture; the addition of cell growth liquid is 900ml, including 738ml basal medium, 72ml Galactose, 90ml grouper tissue fluid; the basal medium is DMEM basal medium, and the preparation process of the grouper tissue fluid is: add 100ml normal saline to 10g grouper tissue, grind and stir, and centrifuge the supernatant at 10000rpm liquid, after sterilizing and filtering, to obtain; the addition amount is ...

Embodiment 3

[0041] Embodiment 3 uses BC-7L bioreactor suspension to culture grouper cell method

[0042] The method for cultivating grouper cells using BC-7L bioreactor comprises the following steps:

[0043] 1) The grouper GS cells were digested with 0.22% trypsin digestion solution to prepare a cell suspension; the trypsin digestion solution, in terms of mass fraction, included 0.01% ethylenediaminetetraacetic acid and 0.22% trypsin ( 1:250);

[0044] 2) Inoculate the cell suspension prepared in step 1) into the BC-7L bioreactor containing cell growth liquid and microcarriers for dynamic culture; the addition of cell growth liquid is 900ml, including 738ml basal medium, 72ml Galactose, 90ml grouper tissue fluid; the basal medium is DMEM basal medium, and the preparation process of the grouper tissue fluid is: add 100ml normal saline to 10g grouper tissue, grind and stir, and centrifuge the supernatant at 10000rpm liquid, after sterilizing and filtering, to obtain; the microcarrier is Cy...

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Abstract

The invention belongs to the technical field of biology, and particularly relates to a method for culturing grouper cells by using a bioreactor. The method provided by the invention comprises the following steps: culturing grouper cells on a large scale by virtue of microcarrier adherent suspension in a BC-7L bioreactor, and adding a GS cell growth solution, namely adding 50-80 ml of galactose and 100-160 ml of grouper tissue culture solution into a DMEM basic culture medium, so that metabolism of lower organisms can be promoted. According to the method provided by the invention, large-scale microcarrier culture of the grouper cells is realized, the yield of the microcarrier cells is high, the cultured cells are normal in morphology, the activity and hereditary characters of original cells are maintained, and the grouper cells have the capabilities of infecting and reproducing viruses and can be used for reproducing fish viruses and producing fish vaccines on a large scale.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for cultivating grouper cells using a bioreactor. Background technique [0002] Bioreactor refers to the equipment that provides a suitable environment for the proliferation of living cells and the biochemical reactions inside the cells. It is the key equipment in the process of biopharmaceuticals. There are many factors that affect the efficiency or function of a bioreactor. The structure of the reactor, the quality of the reactor, the mode of operation, and the selection of operating conditions are closely related to the quality of biological products, production efficiency, and energy consumption. In recent years, bioreactors have been widely used in animal cultivation in the field of biopharmaceuticals, and the total scale of animal cell culture reactors is still developing in a large-scale direction. [0003] The use of bioreactors to cultivate animal cells...

Claims

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Application Information

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IPC IPC(8): C12N5/078
CPCC12N5/0648C12N2509/00C12N2500/34C12N2500/80C12N2531/00
Inventor 尹顺义袁春燕伍活镰蒋东阳任政华
Owner GUANGZHOU QIZHI BIOLOGICAL ENG EQUIP
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