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Stem cell three-dimensional odontogenic induction differentiation method and application

A technology for inducing differentiation and stem cells, applied in the direction of non-embryonic pluripotent stem cells, biochemical equipment and methods, embryonic cells, etc., can solve problems such as the inability to reproduce the three-dimensional microenvironment of cells, the negative effects of stem cell proliferation and differentiation, and inconsistent test results. Achieve the effect of improving reliability and repeatability, simple and easy to manufacture, and low equipment requirements

Pending Publication Date: 2021-12-21
STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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Problems solved by technology

[0004] At present, the main induction culture method of stem cells is mostly two-dimensional culture, which only supports the growth and differentiation of stem cells on one plane, and cannot reproduce the real three-dimensional microenvironment of cells in vivo
The two-dimensional culture environment is far inferior to the three-dimensional culture in many aspects such as biological activity and nutrient release, so that the stem cells gradually lose their original traits, shape, structure and function, resulting in often inconsistent research results and in vivo test results. The culture environment provided is very different from the microenvironment in vivo, which will inevitably have a negative effect on the proliferation and differentiation of stem cells

Method used

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  • Stem cell three-dimensional odontogenic induction differentiation method and application
  • Stem cell three-dimensional odontogenic induction differentiation method and application
  • Stem cell three-dimensional odontogenic induction differentiation method and application

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preparation example Construction

[0029] 2. Preparation of incompletely decalcified dentin matrix:

[0030] Incisors or molars are extracted from the jaws of freshly sacrificed animals, or premolars extracted from orthodontic patients are collected clinically, and the high-speed turbine cuts off the crown part and retains the root part, and then removes all non-dentine tissue and the inner and outer surface parts of the root Dentin, to obtain a dentin matrix. Dentin matrix using ddH 2 O rinsed 3 times, then soaked in ddH 2 O placed in an ultrasonic oscillator cleaner for 20 minutes. Then the cleaned material was placed in 15% EDTA solution for 20-30 minutes, 10% EDTA for 20-30 minutes, and finally in 5% EDTA for 20-30 minutes, ddH 2 O was rinsed for 10 minutes to obtain incompletely decalcified dentin matrix material, which was sterilized with antibiotics or alcohol for later use.

[0031] 3. Three-dimensional odontogenic differentiation of dental pulp stem cells

[0032] The dental pulp stem cells were d...

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Abstract

The invention provides a stem cell three-dimensional odontogenic induction differentiation differentiation method and an application. The method is implemented by the following technical scheme: performing decalcification treatment on human or animal dentin matrixes to open dentin tubules of the human or animal dentin matrixes so as to facilitate release of odontogenic induction factors, putting the dentin matrixes into a low-adsorption culture dish, inoculating a proper amount of stem cells, performing inoculating with an appropriate amount of stem cells, and after culturing for a certain time, inducing cell adhesion and differentiation. A cell three-dimensional induction differentiation culture material comprises dentin matrixes of different species sources, such as human, pig, cattle and the like, and also comprises dentin matrixes with different decalcification degrees. The low-adsorption culture dish is not limited to culture dishes produced by specific companies and other technologies for preventing stem cells from adhering to the dish bottom are also included. The stem cells are not limited to odontogenic stem cells, and also include embryonic stem cells, umbilical cord stem cells, induced pluripotent stem cells, mesenchymal stem cells and the like.

Description

technical field [0001] The invention relates to the field of dental regeneration, in particular to a method and application for inducing three-dimensional odontogenic differentiation of stem cells. Background technique [0002] Tooth loss caused by periodontitis and other diseases and trauma is one of the common and frequently-occurring diseases of human beings, which seriously affects the patient's mastication, speech, aesthetics and mental health. The current treatment methods for missing teeth include removable dentures, fixed dentures and dental implants. These three methods are non-biological repair methods, which have many disadvantages, such as limited recovery of physiological functions, short service life and other defects, and the inability to achieve true physiological functional repair. The use of stem cells and tissue engineering technology to achieve tooth regeneration has become a hotspot in the current international research of stomatology, providing a new s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/0735C12N5/074C12N5/0775
CPCC12N5/0654C12N2533/90C12N2506/02C12N2506/1369C12N2506/45C12N2506/1346C12N2513/00
Inventor 李杰袁瑜
Owner STOMATOLOGICAL HOSPITAL OF CHONGQING MEDICAL UNIV
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