Preparation method of dextral hydrogel material

A gel material, dextro-gel technology, applied in biochemical equipment and methods, microorganisms, bone/connective tissue cells, etc., can solve problems such as difficult to precisely regulate the adipogenic differentiation of stem cells, and achieve the promotion of bone marrow mesenchymal stem cells The effect of adipogenic differentiation

Inactive Publication Date: 2019-02-15
PEKING UNIV SCHOOL OF STOMATOLOGY +1
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the technical problem that the existing matrix materials are difficult to accurately regulate the adipogenic differentiation of stem cells, the present invention provides a preparation method of a dextro-hydrogel material that utilizes the interaction between cells and materials to regulate the adipogenic differentiation of stem cells

Method used

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  • Preparation method of dextral hydrogel material
  • Preparation method of dextral hydrogel material
  • Preparation method of dextral hydrogel material

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Use dextrogel factor dissolved in dimethyl sulfoxide solution to obtain a solution of dextrogel factor with a mass volume concentration of 12mg / ul, and place it at the bottom of a 24-well plate, 500ul containing 100,000 bone marrow mesenchymal stem cells The culture medium suspension was quickly injected into the above-mentioned 24-well plate wells, and allowed to stand at 40°C for 60 minutes to form a dextrorotatory hydrogel.

[0020] Add the dextrorotatory hydrogel to the mesenchymal stem cell medium without osteoinductive factors (the components are mesenchymal stem cell basal medium + 10% fetal bovine serum + 100IU / mL penicillin-streptomycin, all purchased at Saiye Biotechnology Co., Ltd., the same below), during the culture period, the mesenchymal stem cell medium on the D-hydrogel was replaced every 2 days.

[0021] The D-hydrogel mixed with mesenchymal stem cells was cultured for 3 days, and implanted into the subcutaneous area of ​​rats.

[0022] After culturin...

Embodiment 2

[0024] Use dextrogel factor dissolved in dimethyl sulfoxide solution to obtain a solution of dextrogel factor with a mass volume concentration of 23mg / ul, and place it at the bottom of a 24-well plate, 500ul containing 100,000 bone marrow mesenchymal stem cells The culture medium suspension was quickly injected into the above-mentioned 24-well plate wells, and allowed to stand at 35°C for 45 minutes to form a dextrorotatory hydrogel.

[0025] Add the right-handed hydrogel to the mesenchymal stem cell medium without osteoinductive factors, and replace the mesenchymal stem cell medium on the right-handed hydrogel every 2 days during the culture period.

[0026] The D-hydrogel mixed with mesenchymal stem cells was cultured for 3 days, and implanted into the subcutaneous area of ​​rats.

[0027] After culturing the mesenchymal stem cells mixed with the dextrorotatory hydrogel for 7 days, the adipogenic differentiation of bone marrow mesenchymal stem cells was observed by immunoflu...

Embodiment 3

[0029] Use dextrogel factor dissolved in dimethyl sulfoxide solution to obtain a solution of dextrogel factor with a mass volume concentration of 33mg / ul, and place it at the bottom of a 24-well plate, 500ul containing 100,000 bone marrow mesenchymal stem cells The culture medium suspension was quickly injected into the above-mentioned 24-well plate wells, and allowed to stand at 30°C for 30 minutes to form a dextrorotatory hydrogel.

[0030] The D-hydrogel was added to the mesenchymal stem cell culture medium without osteoinductive factors, wherein the mesenchymal stem cell culture medium without osteoinductive factors, during the culture, the interstitial cells on the D-hydrogel were replaced every 2 days. Mesenchymal stem cell culture medium.

[0031] The D-hydrogel mixed with mesenchymal stem cells was cultured for 3 days, and implanted into the subcutaneous area of ​​rats.

[0032] After culturing the mesenchymal stem cells mixed with the dextrorotatory hydrogel for 7 da...

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Abstract

The invention relates to a preparation method of a dextral hydrogel material, and solves the technical problem that an existing matrix material can hardly precisely regulate adipogenic differentiationof stem cells. The preparation method comprises mainly the following main steps: (1) dissolving a dextral gel factor in a dimethyl sulfoxide solution to obtain a dextral gel factor solution with a volumetric concentration of 12 to 33 mg/[mu]l, and placing the solution at the bottom of a 24-pore plate; (2) uniformly mixing the prepared dextral gel factor solution with a culture medium suspension of mesenchymal stem cells in the 24-pore plate, and standing still at 30 to 40 DEG C for 30 to 60 minutes to form dextral hydrogel; (3) putting the prepared dextral hydrogel into an osteoinductive factor-free culture medium suspension of mesenchymal stem cells for culturing, and replacing the culture medium of the mesenchymal stem cells on the dextral hydrogel at intervals. The preparation method can be widely applied to the field of hydrogel fibers for regulating and controlling the adipogenic differentiation of the three-dimensional mesenchymal stem cells.

Description

technical field [0001] The invention relates to the field of biochemical industry, in particular to a preparation method of a dextrorotatory hydrogel material. Background technique [0002] The growth-local microenvironment of stem cells can regulate cell fate and behavior and guide developmental processes. During embryonic development, the extracellular matrix microenvironment is involved in the regulation of embryonic formation and organogenesis. The physical environment of pluripotent stem cells regulates their self-renewal and differentiation. Mechanical and physical cues are also important in adult tissues, where adult stem cells require physical interaction with the extracellular matrix to maintain their potency. Therefore, how to regulate the differentiation and function of stem cells through the extracellular matrix has become a research hotspot in regenerative medicine. Designing and constructing highly bioactive scaffold materials from the perspective of bionics...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077
CPCC12N5/0653C12N2506/1353C12N2513/00C12N2533/00
Inventor 卫彦邓旭亮冯传良江圣杰司梦婷
Owner PEKING UNIV SCHOOL OF STOMATOLOGY
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