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A culture method for maintaining the self-renewal state of mouse epiblast stem cells

A technology for updating state and culture method, applied in the direction of cell culture active agent, cell culture support/coating, embryo cells, etc., which can solve the problems of high cost of culture conditions, unfavorable follow-up exploration, and inconvenient passaging operation.

Active Publication Date: 2019-08-16
ANHUI UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to solve the problems of unstable cell state, inconvenient subculture operation, high cost of existing culture conditions, complex action factors and unfavorable follow-up exploration in traditional culture conditions, the present invention provides a cost-effective, convenient and stable effect. , the culture conditions of mouse epiblast stem cells that are conducive to the development of later research

Method used

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  • A culture method for maintaining the self-renewal state of mouse epiblast stem cells
  • A culture method for maintaining the self-renewal state of mouse epiblast stem cells
  • A culture method for maintaining the self-renewal state of mouse epiblast stem cells

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Experimental program
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Effect test

Embodiment

[0094] (1) Acquisition of mouse epiblast stem cells

[0095] 1) One female and one male mouse of the 129 strain with reproductive ability are caged together, and the formation of the vaginal plug in the vagina of the female mouse is checked before 9 o'clock every morning. When it is first seen, it is recorded as the 0.5th day of mating completion ;

[0096] 2) On the 5.5th day after the completion of mating, the embryo in the uterus of the mother mouse was obtained by dissection, and the epiblast tissue was obtained by separating the extraembryonic tissue with a glass needle;

[0097] 3) Add 1ml of 0.1% gelatin into a Thermo NUNC 4-well cell culture plate, place at 37°C, 5% CO 2 Concentration in the cell culture box, coated for 1h;

[0098] 4) Abandon the gelatin, add 0.5ml DMEM medium to the culture well, which contains: 10% FBS, 1% MEM non-essential amino acids, 2mM L-Glutamin, 0.1mM β-mercaptoethanol, 1mM sodium pyruvate, 100 units of penicillin and 100 μg of streptomyci...

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Abstract

The invention discloses a culture method capable of maintaining the self-renewal state of mouse epiblast stem cells. The culture method includes: acquiring, performing passage on and culturing the mouse epiblast stem cells under culture conditions, and observing and detecting the self-renewal state of the cultured mouse epiblast stem cells. The culture method has the advantages that the problems that the state of cells cultured under traditional culture conditions is unstable, passage operation is inconvenient, and existing culture conditions are high in cost, complex in action factors and unfavorable for subsequent researches, and the like are solved; the method is economic, efficient, convenient to operate, stable in effect, beneficial to researches, and the like; the method can provide a clue for the improvement of the culture conditions of current human embryonic stem cells and other kinds of pluripotent stem cells and plays an important role in the promotion of the smooth development of stem cell foundation and application researches.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to a culture method for maintaining the self-renewal state of mouse epiblast stem cells. Background technique [0002] Stem cells are a type of cells isolated from early embryos. Under in vitro culture conditions, they have the ability to proliferate indefinitely (self-renewal), while maintaining the potential to differentiate into various types of cells in the body (differentiation). It is one of the powerful tools for screening drugs and making animal models of diseases, and has great application prospects in the field of regenerative medicine. [0003] The most widely studied stem cells are mouse Embryonic StemCells (mESCs), which is also the first stem cell line successfully established in vitro. Subsequently, human embryonic stem cell (humanEmbryonic Stem Cells, hESCs) lines were also successfully constructed. Although mouse and human ESCs share some common features, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0735
CPCC12N5/0606C12N2501/999C12N2533/54
Inventor 叶守东孙元元
Owner ANHUI UNIVERSITY
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