Tsingtao amphioxus cell line culture fluid and preparation method thereof

A culture medium and cell line technology, applied in the field of cell biology, can solve problems such as inability to subculture amphioxus cells in vitro

Inactive Publication Date: 2012-10-31
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] The purpose of the present invention is to solve the technical problem that the in vitro subculture of amphioxus cells cannot be achieved in the prior art, provide a stable Qingdao amphioxus cell line and a method for establishing this cell line, and establish a good foundation for the research of amphioxus Experimental platform to facilitate further in-depth research on genetics, immunity, evolution, development and genomics

Method used

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  • Tsingtao amphioxus cell line culture fluid and preparation method thereof
  • Tsingtao amphioxus cell line culture fluid and preparation method thereof
  • Tsingtao amphioxus cell line culture fluid and preparation method thereof

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Experimental program
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Embodiment 1

[0038] The specific steps of establishing the Qingdao amphioxus cell line provided by the present invention are as follows:

[0039] 1) Cultivate primary cells from the gills of Qingdao amphioxus: put Qingdao amphioxus in an ice bath until comatose, disinfect the body surface with 70-75% alcohol, dissect aseptically, take the gill tissue of amphioxus, and use cells containing penicillin and streptomycin After rinsing with the culture medium, cut it into small tissue pieces, transfer it to a six-well cell culture plate, spread it evenly, and culture it at a constant temperature at 25-29°C to make it adhere to the wall. Observe the growth status of the cells every day, and add 0.5-1ml cell culture medium; replace 1 / 2 volume of cell culture medium after 3 days. Such as figure 1 As shown, after the primary culture of amphioxus gill tissue, adherent cells and suspension cells can be seen to dissociate from the tissue. The suspension cells are round and the adherent cells are polyg...

Embodiment 2～5

[0045] The culture medium component that is specially used in the above-mentioned method for establishing Qingdao amphioxus cell line provided by the present invention is preferred:

[0046]

[0047]

[0048]

[0049] Preparation Example

[0050] The preparation provided by the invention is specially used to establish the cell culture fluid of described Qingdao amphioxus cell line method:

[0051] Add the components shown in the table into 0.8 liters of ultrapure water (resistivity 18.2Ω), adjust the osmotic pressure to 750-800mOsm / kg, adjust the pH to 7.0-7.2, and use ultrapure water (resistivity 18.2Ω ) Adjust the total volume of the culture solution to 1 liter, mix well, store at 4°C after filter sterilization; add 10% fetal bovine serum before use.

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Abstract

The invention belongs to the field of cytobiology. The successful subculture of Tsingtao amphioxus gill primary cells is implemented by cDNA transfection of human telomere enzyme catalysis subunit hTERT, thereby establishing the Tsingtao amphioxus cell line. The invention provides a cell culture fluid for cell line establishment, which uses a Leiboviz's L15 culture medium as the basal culture medium and contains the following components: at least one essential amino acid and non-essential amino acid, at least one vitamin, at least one salt, at least one buffer solution and saccharide, and at least one phenolic compound. The invention also provides a method for preparing the culture fluid. The invention successfully solves the technical problems in in-vitro subculture of amphioxus cells, and is beneficial to deep research in the fields of genetics, immunology, evolution, auxology, genomics and the like.

Description

[0001] This application is a divisional application of a Chinese invention patent application with the application date of December 29, 2010, the application number 201010612800.0, the applicant is Sun Yat-sen University, and the invention name is "A Qingdao amphioxus cell line and its establishment method". technical field [0002] The invention belongs to the field of cell biology, and in particular relates to a Qingdao amphioxus cell line culture fluid and a method for preparing the culture fluid. Background technique [0003] The marine model animal Amphioxus, known in Latin literature as Branchiostoma belcheri, belongs to the primitive cord animal. It not only retains some characteristics of invertebrates, but also has the characteristics of vertebrates. It is a transitional type between invertebrates and vertebrates. Amphioxus appeared as early as 500 million years ago, and is considered to be the representative of the ancestor of the existing vertebrates, known as the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10
Inventor 张雁王华敏蔡晓青
Owner SUN YAT SEN UNIV
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