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Aminated graphene culture mediumas well as preparation method and application thereof in regulation of induced pluripotent stem cells

A technology of aminated graphene and pluripotent stem cells, applied in the field of biomedical engineering, can solve the problems of immature cardiomyocytes being difficult to apply in vivo, immature cardiomyocytes, etc., achieving good biocompatibility, improving remodeling effect, Prepare simple effects

Active Publication Date: 2020-11-10
HUAZHONG UNIV OF SCI & TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The invention solves the technical problem in the prior art that the differentiation of induced pluripotent stem cells into cardiomyocytes is not mature enough, which makes immature cardiomyocytes difficult to apply in vivo

Method used

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  • Aminated graphene culture mediumas well as preparation method and application thereof in regulation of induced pluripotent stem cells
  • Aminated graphene culture mediumas well as preparation method and application thereof in regulation of induced pluripotent stem cells
  • Aminated graphene culture mediumas well as preparation method and application thereof in regulation of induced pluripotent stem cells

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Experimental program
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Effect test

Embodiment 1

[0034] Dissolve 10 mg of aminated graphene in 10 ml of ultrapure water, and use ultrasonic treatment to uniformly disperse to obtain a 1 mg / ml aminated graphene dispersion. Aspirate 30 μl of the dispersion solution for vacuum filtration onto a microporous membrane, followed by drying at room temperature. Squeeze the filter membrane with the aminated graphene film side on the cover glass, squeeze it by 20N gravity for 10 hours, transfer the aminated graphene film to the cover glass, and dissolve the microporous filter film in acetone for 2 hours, wash with methanol After 10 minutes of treatment, the aminated graphene culture substrate can be obtained.

Embodiment 2

[0036] Dissolve 5 mg of aminated graphene in 10 ml of ultrapure water, and use ultrasonic treatment to uniformly disperse to obtain a 0.5 mg / ml aminated graphene dispersion. Aspirate 30 μl of the dispersion solution for vacuum filtration onto a microporous membrane, followed by drying at room temperature. Squeeze the filter membrane with the aminated graphene film side on the cover glass, squeeze the aminated graphene film for 8 hours by 10N gravity, transfer the aminated graphene film to the cover glass, dissolve the microporous filter film in acetone for 2 hours, and wash with methanol After 10 minutes of treatment, the aminated graphene culture substrate can be obtained.

Embodiment 3

[0038] Dissolve 20 mg of aminated graphene in 10 ml of ultrapure water, and use ultrasonic treatment to uniformly disperse to obtain a 2 mg / ml aminated graphene dispersion. Aspirate 30 μl of the dispersion solution for vacuum filtration onto a microporous membrane, followed by drying at room temperature. Squeeze the filter membrane with the aminated graphene film side on the cover glass, squeeze the aminated graphene film for 12 hours by 50N gravity, transfer the aminated graphene film to the gelatin film, and dissolve the microporous filter membrane with ethyl acetate for 2 hours, After washing with methanol for 10 minutes, the aminated graphene culture substrate can be obtained.

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Abstract

The invention discloses an aminated graphene culture medium as well as a preparation method and an application thereof in regulation of induced pluripotent stem cells, and belongs to the field of biomedical engineering. The preparation method comprises the following steps: dissolving aminated graphene in water, and uniformly dispersing to obtain aminated graphene dispersion liquid; carrying out vacuum filtration on the aminated graphene dispersion liquid to the surface of a microporous filter membrane, and performing drying to form an aminated graphene membrane loaded on the surface of the microporous filter membrane; carrying out gravity extrusion on the microporous filter membrane to transfer the aminated graphene membrane to a support layer, namely attaching the aminated graphene membrane to the surface of the support layer; and dissolving the microporous filter membrane, washing off the residual solvent, and performing drying to obtain the aminated graphene culture medium. The culture medium provided by the invention has a unique positively charged amino group, regulates the directional myocardial differentiation of the induced pluripotent stem cells (iPS), and can effectivelystimulate maturation of myocardial precursor cells.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and more specifically relates to an aminated graphene culture substrate and the application of preparing and regulating induced pluripotent stem cells. Background technique [0002] Heart disease remains the leading cause of death worldwide, and infarcted areas of injured hearts are difficult to restore due to a lack of mature cardiomyocytes. The emergence of induced pluripotent stem cells provides a possible source of cells, which have a high degree of similarity to embryonic stem cells in terms of morphology, self-renewal and pluripotency, but there are no ethical issues. [0003] From the initial pluripotent stage to the final differentiated cells of induced pluripotent stem cells, culture substrates play a crucial role in their fate. At present, the most commonly used substrate for the maintenance and differentiation of induced pluripotent stem cells is the substrate-coated culture dish...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/074C01B32/194
CPCC12N5/0657C01B32/194C12N2506/45C12N2533/20C12N2511/00
Inventor 王江林许银
Owner HUAZHONG UNIV OF SCI & TECH
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