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Preparation method of L-hydrogel material

A technology of hydrogel and production method, which is applied in the fields of medical science, tissue regeneration, prosthesis, etc., can solve problems such as the difficulty in precisely regulating the osteogenic differentiation of stem cells, and achieve the promotion of osteogenic differentiation of bone marrow mesenchymal stem cells and obvious bone repair effect of effect

Active Publication Date: 2019-02-12
PEKING UNIV SCHOOL OF STOMATOLOGY +1
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  • Abstract
  • Description
  • Claims
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Problems solved by technology

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

Method used

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  • Preparation method of L-hydrogel material
  • Preparation method of L-hydrogel material
  • Preparation method of L-hydrogel material

Examples

Experimental program
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Effect test

Embodiment 1

[0022] Use levogel factor dissolved in dimethyl sulfoxide solution to obtain a levo-gel factor solution with a mass volume concentration of 33mg / ul, and place it at the bottom of a 24-well plate, 500ul of a medium containing 100,000 bone marrow mesenchymal stem cells The suspension was quickly injected into the wells of the above-mentioned 24-well plate, and allowed to stand at 40° C. for 60 minutes to form a levorotatory hydrogel.

[0023] Add the left-handed 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, both purchased from Sai Industry Biotechnology Co., Ltd., the same below), during the culture period, the mesenchymal stem cell culture medium on the L-hydrogel was replaced every 2 days.

[0024] After the mesenchymal stem cells were mixed with levorotatory hydrogel and cultured for 7 days, bone marrow mesenchymal stem cells were d...

Embodiment 2

[0026] Dissolve the left-handed gelatin factor in dimethyl sulfoxide solution to obtain a solution with a mass volume concentration of 23 mg / ul left-handed gelatin factor, and place it at the bottom of a 24-well plate, culture 500ul of 100,000 bone marrow mesenchymal stem cells The base suspension was quickly injected into the wells of the above-mentioned 24-well plate, and allowed to stand at 35° C. for 45 minutes to form a levorotatory hydrogel.

[0027] Add the left-handed hydrogel to the mesenchymal stem cell medium without osteoinductive factors, and replace the mesenchymal stem cell medium on the left-handed hydrogel every 2 days during the culture period.

[0028] After the mesenchymal stem cells mixed with levorotatory hydrogel were cultured for 7 days, the osteogenic differentiation of bone marrow mesenchymal stem cells was detected by immunofluorescence.

Embodiment 3

[0030] Use the left-handed gel factor dissolved in dimethyl sulfoxide solution to obtain the left-handed gel factor solution with a mass volume concentration of 12mg / ul, and place it at the bottom of a 24-well plate, and culture 500ul of 100,000 bone marrow mesenchymal stem cells The base suspension was quickly injected into the wells of the above-mentioned 24-well plate, and allowed to stand at 30° C. for 30 minutes to form a levorotatory hydrogel.

[0031] The left-handed hydrogel was added to the mesenchymal stem cell culture medium without osteoinductive factors, wherein the mesenchymal stem cell culture medium without osteogenic inductive factors, during the culture period, the mesenchyme on the left-handed hydrogel was replaced every 2 days Stem Cell Medium.

[0032] After cultured for 3 days with levorotatory hydrogel mixed with mesenchymal stem cells, they were implanted into the surgical area of ​​the rat skull defect model. Eight weeks later, the microCT of the skull...

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Abstract

The invention relates to a preparation method of an L-hydrogel material, which solves the technical problem of difficulty in precise regulating and controlling of osteogenesis differentiation of stemcells in the existing substrate material. The preparation method mainly comprises the following steps of (1) dissolving L-gel factors into a dimethyl sulfoxide solution to obtain an L-gel factor solution with mass volume concentration of 12 to 33 mg / ul, and putting the L-gel factor solution at the bottom of a 24-pore plate; (2) mixing the prepared L-gel factor solution into culture medium suspension liquid of mesenchymal stem cells, uniformly mixing in the 24-pore plate, standing for 30 to 60 min at a temperature of 30 to 40 DEG C, and forming a hydrogel; (3) putting the prepared hydrogel intoa mesenchymal stem cell culture medium without osteogenesis inducing factors to culture, and replacing the mesenchymal stem cell culture medium at intervals. The preparation method can be widely applied to the field of regulating and controlling the osteogenesis differentiation of three-dimensional mesenchymal stem cells by hydrogel fiber.

Description

technical field [0001] The invention relates to the field of hydrogel biological implant materials, in particular to a preparation method of a left-handed 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 perspect...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/52A61L27/54C12N5/0775
CPCA61L27/3834A61L27/3847A61L27/3895A61L27/52A61L27/54A61L2430/02C12N5/0663
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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