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A conductive extracellular matrix composite film obtained through cell sheets and its preparation method

A technology of composite film and external matrix, applied in medical science, prosthesis, etc., can solve problems affecting functionality, etc., and achieve good electrical conductivity, easy implementation, good biocompatibility and electrical conductivity

Active Publication Date: 2020-05-29
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, pure mechanical peeling will cause certain changes in the ultrastructure of the extracellular matrix and affect its subsequent functional properties.

Method used

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  • A conductive extracellular matrix composite film obtained through cell sheets and its preparation method
  • A conductive extracellular matrix composite film obtained through cell sheets and its preparation method
  • A conductive extracellular matrix composite film obtained through cell sheets and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] (1) For coated TiO 2 The surface of the nano-dot film is sterilized;

[0025] (2) Detach the cells previously cultured in the culture flask, centrifuge them at 800r / min for 6min, suspend them with α-MEM medium, and count them with a hemocytometer at 1×10 5 piece / cm 2 Cell density Seed the cells on the cell culture surface, place them in a carbon dioxide incubator for high-density culture for 10 days, change the medium for the first time after 2 days, then change the medium every 2 days, and add 100 μg of it one day before cell desorption / mL graphene nanosheets, eventually forming a complete cell sheet on the cell culture surface;

[0026] (3) By irradiating with ultraviolet light with a wavelength of 365nm for 30min, the cultured cell sheet containing conductive substances can be desorbed to obtain a complete cell sheet, and the desorbed cell sheet can be detached with PBS buffer and deionized water. The cell sheet was washed twice repeatedly.

[0027] (4) Soak the...

Embodiment 2

[0030] (1) For coated TiO 2 The surface of the nano-dot film is sterilized;

[0031] (2) Detach the cells previously cultured in the culture flask, centrifuge at 1300r / min for 2min, suspend with α-MEM medium, and count with a hemocytometer at 2×10 6 piece / cm 2 Cells were inoculated on the surface of the cell culture, and placed in a carbon dioxide incubator for high-density culture for 7 days. After 2 days, the medium was changed for the first time, and the medium was changed every 2 days thereafter. 1 μg was added 3 days before cell desorption. / mL graphene nanodots, eventually forming a complete cell sheet on the cell culture surface;

[0032] (3) By irradiating with ultraviolet light with a wavelength of 365nm for 5 minutes, the cultured cell sheet containing conductive substances can be desorbed to obtain a complete cell sheet, and the desorbed cell sheet can be detached with PBS buffer and deionized water. The cell sheet was washed 3 times repeatedly.

[0033] (4) Soa...

Embodiment 3

[0036] (1) For coated TiO 2 The surface of the nano-dot film is sterilized;

[0037] (2) Detach the cells previously cultured in the culture flask, centrifuge them at 1000r / min for 4min, suspend them with α-MEM medium, and count them with a hemocytometer at 5×10 4 piece / cm 2 Cells were inoculated on the surface of the cell culture, and placed in a carbon dioxide incubator for high-density culture for 15 days. After 2 days, the medium was changed for the first time, and the medium was changed every 3 days thereafter. 10 μg was added 2 days before cell desorption. / mL carbon nanofibers, and finally form a complete cell sheet on the cell culture surface;

[0038] (3) By irradiating with ultraviolet light with a wavelength of 365nm for 15 minutes, the cultured cell sheet containing conductive substances can be desorbed to obtain a complete cell sheet, and the desorbed cell sheet can be detached with PBS buffer and deionized water. The cell sheet was washed 3 times repeatedly. ...

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Abstract

The invention discloses a conductive extracellular matrix composite film acquired from a cell sheet and a preparation method of the composite film. The preparation method includes the steps: culturing cells on the cell culture surface (a TiO2 nano-dot film) capable of photocell detachment, adding nano-conductive materials in the culturing process, acquiring a complete cell sheet through the photocell detachment after fusion of the cell sheet, cleaning the cell sheet, and performing decellularization on the cell sheet to obtain the conductive extracellular matrix composite film. The nano-conductive materials include a graphene nano-sheet, a graphene nano-point and the like, nano-conductive particles can be internally swallowed by cells, and the conductive materials are dispersed in the cells and between cells. The conductive extracellular matrix composite film prepared by the method can be used for the field of tissue engineering and the like. The preparation method is simple in process and easy to realize, and popularization and application are facilitated.

Description

technical field [0001] The invention relates to the field of tissue engineering, in particular to a conductive extracellular matrix composite film obtained through cell sheets and a preparation method thereof. Background technique [0002] The extracellular matrix is ​​one of the basic components of tissues. It is composed of some glycoproteins, structural proteins and adhesive proteins. It can carry out all-round biological regulation on the basic life activities of cells, such as the shape, growth, migration and differentiation of cells. , the development of embryos and the repair of damaged tissues or organs have a vital role. Studies have reported that the synergistic effect of simulated extracellular matrix environment such as collagen and hyaluronic acid and electrical stimulation can effectively promote the osteogenic differentiation of human bone marrow mesenchymal stem cells [Hess R, Jaeschke A, Neubert H, et al. Synergistic effect of defined artificial extracellul...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/56A61L27/58A61L27/12A61L27/50A61L27/36
CPCA61L27/12A61L27/3633A61L27/3891A61L27/3895A61L27/50A61L27/56A61L27/58
Inventor 程逵王小召陈樽翁文剑
Owner ZHEJIANG UNIV
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