Improved L-15 culture medium and application thereof in culturing prawn muscle cells

A muscle cell and culture medium technology, applied in the field of improved L-15 medium and its application in cultivating prawn muscle cells, can solve the problems of culturing for hours or even days, lack of accurate determination of amino acid requirements, tissue blocks The culture method is susceptible to pollution and other problems, so as to reduce the risk of contamination, improve the growth state of cells, and increase the survival rate and adhesion rate

Active Publication Date: 2021-06-25
SOUTH CHINA NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the above-mentioned patent does not accurately determine the amino acid requirements of the cells, and only expands all components of the L-15 medium by 1.5 times
Moreover, the primary cell extraction method is tissue block culture, which requires several hours or even days of culture to obtain cells, and the tissue block culture method is more likely to be contaminated during the culture process
The medium working solution of this patent also added 20ng / mL basic fibroblast growth factor (bFGF) and 20ng / mL epidermal growth factor (EGF), which greatly increased the cost

Method used

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  • Improved L-15 culture medium and application thereof in culturing prawn muscle cells
  • Improved L-15 culture medium and application thereof in culturing prawn muscle cells
  • Improved L-15 culture medium and application thereof in culturing prawn muscle cells

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Experimental program
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Effect test

Embodiment 1

[0051] Prepare the prawn muscle cell extract, comprising the following steps:

[0052] (1) Sterilize the Penaeus vannamei, then cut the muscle, put it into the PBS solution containing 5% double antibodies (penicillin and streptomycin; the same below) to rinse, then put it into the ethanol solution to rinse, and then put it into Rinse in PBS solution containing 5% double-antibody, and finally soak in L-15 medium containing 5% double-antibody for at least 30 minutes;

[0053](2) Take out the prawn muscles and cut them into chylus, add L-15 medium containing 10% fetal bovine serum (FBS) and 1% double antibody, blow the prawn muscles evenly, pass through a 100μm cell sieve, and then add The L-15 medium containing 5% double antibody was resuspended and centrifuged, and finally diluted with L-15 medium containing 10% fetal bovine serum (FBS) and 1% double antibody to obtain the prawn muscle cell extract;

[0054] The muscle cells of Penaeus vannamei were inoculated in a 96-well pla...

Embodiment 2

[0076] According to the experimental results of Example 1, the present invention designs a kind of improved L-15 culture medium, contains 10mmol / L alanine, 3mmol / L glycine, 3mmol / L serine, 1.5mmol / L histidine, 0.5mmol / L Lysine, 1.5mmol / L arginine, 1.5mmol / L valine, 1mmol / L threonine, 0.5mmol / L methionine, 0.5mmol / L leucine, 5mmol / L aspartic acid, 0.5 mmol / L glutamine, 1mmol / L phenylalanine, the types and concentrations of other raw materials in the medium are the same as those of L-15 medium; it also contains 2.5mmol / L taurine, 50mmol / L proline, 20mmol / L L glutamic acid, 10% FBS, 100 IU / ml penicillin and 0.1 mg / ml streptomycin, pH 7.2-7.4.

[0077] The muscle cells of Penaeus vannamei were inoculated in a 96-well plate and cultured. The initial inoculation of cells was 40% of the area of ​​each well of the culture dish. 100 μL of medium was added to each well and the amount of cells added to each well was the same.

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Abstract

The invention discloses an improved L-15 culture medium and application thereof in culturing prawn muscle cells. The improved L-15 culture medium contains 10 mmol / L of alanine, 3 mmol / L of glycine, 3 mmol / L of serine, 1.5 mmol / L of histidine, 0.5 mmol / L of lysine, 1.5 mmol / L of arginine, 1.5 mmol / L of valine, 1 mmol / L of threonine, 0.5 mmol / L of methionine, 0.5 mmol / L of leucine, 5 mmol / L of aspartic acid, 0.5 mmol / L of glutamine and 1 mmol / L of phenylalanine. The oral liquid also contains taurine, proline, glutamic acid, fetal calf serum and double antibodies, and the pH value of the oral liquid is 7.2-7.4. According to the improved L-15 culture medium, the survival rate and the adherence rate of the primary muscle cells of the penaeus vannamei boone are increased, and the growth state of the cells is improved.

Description

technical field [0001] The invention relates to an improved L-15 medium and its application in culturing prawn muscle cells. Background technique [0002] The establishment of shrimp cell lines is the basis for studying the pathogenic mechanism of viruses, isolating and purifying shrimp viruses, and developing effective diagnostic reagents. The successful establishment of shrimp cell lines can not only precisely control the experimental conditions, but also greatly reduce the experimental cost and obtain better research results. The culture medium is an indispensable part of the cultured cells. The existing culture medium is not enough for the establishment of the shrimp cell line. Therefore, the optimization of the cell culture medium is one of the keys to the successful establishment of the shrimp cell line. This is of great significance for the study of the shrimp cell culture in vitro . [0003] Currently there is no cell culture medium specifically for prawns, and the...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/07
CPCC12N5/0601C12N2500/32C12N2500/33
Inventor 叶超霞邹丹阳
Owner SOUTH CHINA NORMAL UNIVERSITY
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