Aminated graphene culture substrate and its preparation and regulation of induced pluripotent stem cells

A technology of aminated graphene and pluripotent stem cells, which is applied in the field of biomedical engineering, can solve the problems of immature cardiomyocytes that are difficult to apply in vivo, and cardiomyocytes are not mature enough to achieve good biocompatibility, improve remodeling effect, Prepare simple effects

Active Publication Date: 2022-07-05
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 substrate and its preparation and regulation of induced pluripotent stem cells
  • Aminated graphene culture substrate and its preparation and regulation of induced pluripotent stem cells
  • Aminated graphene culture substrate and its preparation and regulation of induced pluripotent stem cells

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Embodiment 1

[0034] Dissolve 10 mg of graphene aminated in 10 ml of ultrapure water, and use ultrasonic treatment for uniform dispersion to obtain 1 mg / ml of graphene aminated dispersion. Aspirate 30 μl of the dispersion liquid, carry out vacuum filtration onto a microporous membrane, and then dry at room temperature. The filter membrane with the aminated graphene film side was squeezed on the cover glass, squeezed by 20N gravity for 10h, the aminated graphene film was transferred to the cover glass, and the microporous filter membrane was dissolved in acetone for 2h, and washed with methanol. After 10 min of treatment, the aminated graphene culture substrate can be obtained.

Embodiment 2

[0036] 5 mg of the aminated graphene was dissolved in 10 ml of ultrapure water, and uniformly dispersed by ultrasonic treatment to obtain 0.5 mg / ml of the aminated graphene dispersion. Aspirate 30 μl of the dispersion liquid, carry out vacuum filtration onto a microporous membrane, and then dry at room temperature. The filter membrane with the graphene aminated film side was extruded on the cover glass, squeezed by 10N gravity for 8 hours, and the graphene aminated film was transferred to the cover glass, and the microporous filter membrane was dissolved in acetone for 2 hours, and washed with methanol. After 10 min of treatment, the aminated graphene culture substrate can be obtained.

Embodiment 3

[0038] Dissolve 20 mg of graphene aminated in 10 ml of ultrapure water, and use ultrasonic treatment for uniform dispersion to obtain a graphene aminated dispersion of 2 mg / ml. Aspirate 30 μl of the dispersion liquid, carry out vacuum filtration onto a microporous membrane, and then dry at room temperature. The filter membrane with the aminated graphene film side was extruded on the cover glass, squeezed by 50N gravity for 12h, the aminated graphene film was transferred to the gelatin film, and the microporous filter membrane was dissolved in ethyl acetate for 2h, After washing with methanol for 10 min, the aminated graphene culture substrate can be obtained.

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Abstract

The invention discloses an aminated graphene culture substrate and the application of preparation and regulation of induced pluripotent stem cells, belonging to the field of biomedical engineering. The preparation method is as follows: dissolving the aminated graphene in water and dispersing it uniformly to obtain the aminated graphene dispersion; vacuum-filtering the aminated graphene dispersion to the surface of a microporous filter membrane, and after drying, the aminated graphene The dispersion liquid forms the aminated graphene film and is loaded on the surface of the microporous filter membrane; the microporous filter membrane is subjected to gravity extrusion, so that the aminated graphene film is transferred to the support layer, even if the aminated graphene film is attached It is attached to the surface of the support layer; the microporous filter membrane is dissolved, the residual solvent is washed away, and after drying, the aminated graphene culture substrate is obtained. The culture substrate provided by the invention has a unique positively charged amino group, regulates the directional myocardial differentiation of induced pluripotent stem cells (iPS), and can effectively stimulate the maturation of myocardial precursor cells.

Description

technical field [0001] The invention belongs to the field of biomedical engineering, and more particularly relates to an aminated graphene culture substrate and an application of preparing and regulating induced pluripotent stem cells. Background technique [0002] Heart disease remains the leading cause of death worldwide, and the infarcted area of ​​an injured heart is difficult to recover due to a lack of mature cardiomyocytes. The emergence of induced pluripotent stem cells provides the possibility of a cell source, which is highly similar to embryonic stem cells in terms of morphology, self-renewal and pluripotency, but there are no ethical issues. [0003] The culture medium plays a crucial role in the fate of induced pluripotent stem cells from the initial pluripotent stage to the final differentiated cell. Currently, the most commonly used matrix for the maintenance and differentiation of induced pluripotent stem cells is the matrix-coated culture dish. However, in...

Claims

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Application Information

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Patent Type & Authority Patents(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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