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Cell micro-patterning method for maintaining stemness of human dental pulp stem cells

A technology of dental pulp stem cells and micropatterns, applied in the field of cell culture, can solve the problems of loss of stemness and differentiation ability, achieve excellent cell function, low production cost, and simple principle

Inactive Publication Date: 2021-04-02
BEIHANG UNIV
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Problems solved by technology

Although many breakthroughs have been made in the research and application of dental pulp stem cells, they are mainly limited to animal models. There are still many problems in order to successfully use them in clinical tissue regeneration. One of the problems is that in vitro culture models, Dental pulp stem cells will gradually lose their stemness and differentiation ability due to continuous renewal and replication, and cannot continue to repair and rebuild periodontal tissue

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  • Cell micro-patterning method for maintaining stemness of human dental pulp stem cells
  • Cell micro-patterning method for maintaining stemness of human dental pulp stem cells
  • Cell micro-patterning method for maintaining stemness of human dental pulp stem cells

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Embodiment Construction

[0023] The cell micropattern technology is simple and easy, and the implementation method of the present invention is as follows: first, take a 0.17 mm glass slide, use an ultraviolet cleaning machine to sterilize its surface, and then use polydimethylsiloxane PDMS in a biological safety cabinet The micropattern stamp modified the fibronectin FN in the above two micropattern patterns on the surface of the slide, and then the slide was modified with poloxamer Pluronic F-127 to isolate the area of ​​the slide that was not modified by fibronectin FN. Then the cell suspension was placed on the glass slide, and after the human dental pulp stem cells adhered to the wall and observed under the microscope that the shape of the cells was completely consistent with the shape of the above-mentioned micro-pattern, the medium was replaced normally for routine culture. The ability to maintain the stemness of human dental pulp stem cells was enhanced, and the differentiation potential of the ...

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Abstract

The invention belongs to the technical field of cell culture methods, and designs a cell micro-pattern method for maintaining stemness of human dental pulp stem cells. The cell micro-pattern method comprises two cell micro-pattern arrangement modes: a rectangular micro-pattern array with the row spacing of 90 microns and the column spacing of 45 microns is formed by 5,000 or more rectangular micro-patterns with the length of 45 microns and the width of 22.5 microns, and a micro-pattern array with the row spacing of 128 microns and the column spacing of 28 microns is formed by 5,000 or more rectangular micro-patterns with the length of 64 microns and the width of 14 microns. The method is used for cell culture inoculation of the human dental pulp stem cells and is simple and easy to operate, cell stemness markers SOX2 and Nanog are dyed through immunofluorescence, it can be proved from expression of the two markers that stemness and differentiation potential of the human dental pulp stem cells can be realized, and the method is a cheap and efficient cell culture method, and is favorable for application of a clinical tissue engineering technology.

Description

technical field [0001] The invention belongs to the technical field of cell culture methods, is based on the principle of tissue engineering, protein microarray technology and cell micropattern technology, and is used to maintain the stemness and differentiation ability of human dental pulp stem cells. Background technique [0002] Periodontitis is a major oral disease that endangers human oral and systemic health, and severe cases can lead to tooth loss. Once adult teeth are lost, the commonly used clinical treatment methods are limited to guided tissue regeneration, bone grafting, etc. Although there are certain curative effects, they still cannot fully restore the function and physiological state of tooth tissue. Therefore, regeneration and reconstruction of functionally meaningful dental tissue is the focus of completely curing periodontitis and even tooth loss. In recent years, with the birth and development of tissue engineering, relying on seed cells for dental tissu...

Claims

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Application Information

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IPC IPC(8): C12N5/0775
CPCC12N5/0663C12N2535/10
Inventor 郑丽沙李驰宇郭雨薇樊瑜波杨智杰宋亚欣纳静石秋生
Owner BEIHANG UNIV
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