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Separation method and culture medium for ricefield eel germline stem cells

A reproductive stem cell and culture medium technology, applied in the field of eel reproductive stem cell separation method and culture medium, can solve the problem of long culture period

Active Publication Date: 2021-01-22
INST OF AQUATIC LIFE ACAD SINICA
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  • Abstract
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  • Application Information

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Problems solved by technology

Xiao Yamei et al. tried to culture rice field eel adult gonad tissue stem cells by separating tissue pieces, but the culture period of this method was long, and the reproductive stem cells obtained were a mixed type of cells.

Method used

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  • Separation method and culture medium for ricefield eel germline stem cells
  • Separation method and culture medium for ricefield eel germline stem cells
  • Separation method and culture medium for ricefield eel germline stem cells

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Embodiment Construction

[0039] The principles and features of the present invention are described below in conjunction with the accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0040] 1. Preparation of eel reproductive stem cell culture medium

[0041] During the isolation and culture of rice field eel reproductive stem cells, the optimization of the medium is very important. After trial and error, we unexpectedly found that, unlike other fish germ stem cells, rice field eel germ stem cells are particularly sensitive to the concentration of FBS. When the concentration of FBS is high, the cultured rice field eel germ stem cells are prone to differentiation, resulting in poor proliferation after attachment . Such as figure 1 As shown, the germinal stem cells of rice field eel with the same concentration were cultured in medium containing 5% and 15% FBS respectively, and the results showed ...

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Abstract

The invention relates to a culture medium for ricefield eel germline stem cells. The culture medium comprises a basic culture medium and FBS, wherein the concentration of the FBS is 5-8%. The invention also relates to a method for separating and culturing the ricefield eel germline stem cells. The method comprises the step of culturing the ricefield eel germline stem cells by using the culture medium. According to the culture medium disclosed by the invention, the adherence efficiency of the ricefield eel germline stem cells can be improved, differentiation of the ricefield eel germline stem cells is prevented, and the proliferation function of the ricefield eel germline stem cells is not influenced. By the method disclosed by the invention, ricefield eel female germline stem cells and male germline stem cells which have higher purity can be separated and obtained in five days by using gonad tissues of ricefield eels. The obtained germline stem cells have typical stem cell morphology,typical stem cell clone can be formed, alkaline phosphatase staining is positive, and related molecular markers of the stem cells are also highly expressed. After the germline stem cells obtained by the method disclosed by the invention are cryopreserved, the germline stem cells can be successfully recovered.

Description

technical field [0001] The invention relates to the field of economic fish breeding, and more particularly relates to a separation method and culture medium for eel reproductive stem cells. Background technique [0002] Germ stem cells are the only seed cells that can transmit genetic material to offspring in the germline maintenance of animals, and have self-renewal function and differentiation pluripotency. The research on reproductive stem cells is very important to animal genetic breeding and conservation of precious species. [0003] The isolation, induced differentiation and related operation techniques of germ stem cells have become hot spots in the field of stem cell research. Although the research on fish stem cells started relatively late, it has made great progress. At present, scholars at home and abroad have successfully established multiple fish embryonic stem cell lines and reproductive stem cell lines. PGCs from rainbow trout and medaka cultured in vitro c...

Claims

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Application Information

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IPC IPC(8): C12N5/075C12N5/076
CPCC12N5/0609C12N5/061C12N2509/00C12N2501/115C12N2501/11C12N2500/30C12N2500/44C12N2501/727Y02A40/81
Inventor 孙玉华孙晓云王永鑫
Owner INST OF AQUATIC LIFE ACAD SINICA
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