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Method for preparing artificial microenvironment and application thereof

An artificial micro-environment technology, applied in the field of biomedical engineering, can solve problems to be improved, and achieve the effects of convenient application, good plasticity, and short production cycle

Active Publication Date: 2016-10-12
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the decellularization matrix materials of artificial synthetic substitute materials and natural tissues still need to be improved

Method used

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  • Method for preparing artificial microenvironment and application thereof
  • Method for preparing artificial microenvironment and application thereof
  • Method for preparing artificial microenvironment and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0082] Example 1. Preparation of three-dimensional porous scaffold and high-throughput scaffold platform by chemical gelation and freeze-drying

[0083] 1. Preparation of three-dimensional porous cell scaffold skeleton

[0084] 1. Laser engraving to prepare the required mold

[0085]A Rayjet laser engraving machine was used to cut polymethyl methacrylate (PMMA) flat plates with thicknesses of 0.3mm, 0.5mm, 1mm, 1.5mm, 2mm, and 3mm (with isolation films attached to both sides) to form mold components. The mold component design is completed by the software AutoCAD, which can be designed into different shapes and sizes according to the needs. For example, in the circular mold, we use a ring with an outer diameter of 5cm and an inner diameter of 2.5cm to realize the circular structure of the pouring material; by changing the shape of the inner ring It can be made into square, triangular and many other shapes of mold components. The main processing parameters of the laser engravi...

Embodiment 2

[0094] Example 2. Functional cells are automatically loaded onto a three-dimensional porous cell scaffold to form a bionic artificial tissue

[0095] The three-dimensional porous cell support of the present invention has good mechanical properties and good water absorption, so cells can be directly absorbed into the support by dropping. In this embodiment, mesenchymal stem cells are taken as an example for illustration.

[0096] 1. Prepare the gelatin three-dimensional porous scaffold according to Example 1.

[0097] 2. Ultraviolet sterilizing the freeze-dried three-dimensional porous scaffold.

[0098] 3. The cells were treated with trypsin and prepared into a single-cell suspension with a certain concentration. Pipette the cell suspension and drop it directly onto the surface of the three-dimensional porous scaffold, and it can be seen that the cell suspension is completely absorbed after a few seconds. Wait 2 hours in a carbon dioxide incubator to allow cells to adhere t...

Embodiment 3

[0100] Example 3. Obtaining free biomimetic microtissues through a high-throughput scaffold platform

[0101] 1. Preparation of special top plate

[0102] The special top plate is made of polydimethylsiloxane (PDMS) by soft etching technology. Take the special top plate corresponding to the high-throughput support platform as an example to prepare: mix PDMS evenly according to the mass ratio of the main agent and the hardener at a ratio of 10:1. The high-throughput mold plate prepared in Example 1 was placed in the pre-made PDMS groove with a depth of 2-3 mm. Pour the bubble-free PDMS mixture into the mold, and ensure that there is a certain thickness of PDMS on the surface of the mold; then place a glass slide on the surface, and then evacuate to the small hole of the mold and there is no gap between the mold and the glass. Air bubbles are present.

[0103] 2. Obtain free three-dimensional porous micro-scaffolds

[0104] After obtaining the prepared miniaturized scaffold ...

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Abstract

The invention relates to a method for preparing an artificial microenvironment and application thereof. Specifically, the method comprises the steps that firstly, a solid-phase carrier is inoculated with functional cells; secondly, the solid-phase carrier obtained in the first step is kept for preset time under the condition which is suitable for growing of the functional cells; thirdly, the solid-phase carrier obtained in the second step is subjected to de-cellularization treatment so that the artificial microenvironment can be obtained. By means of the method, the artificial microenvironment which has the advantages that low immunogenicity and good controllability are achieved, the quality is stable, the production cycle is short and components are controllable and can achieve customization can be prepared.

Description

technical field [0001] The invention relates to the field of biomedical engineering, in particular, the invention relates to a method for preparing an artificial microenvironment and its application. Background technique [0002] The development of regenerative medicine has brought new hope to the complex and serious diseases that are difficult to be treated by drugs. It has gradually become an important direction for the development of clinical medicine and is expected to become the next pillar industry of the medical and health industry after drug and device treatment. At present, regenerative medicine has been successfully used clinically for skin regeneration, articular cartilage reconstruction, tendon and spinal cord injury repair, immune system function reconstruction, etc., and has been used in the treatment of difficult diseases (such as genetic diseases and cardiovascular diseases) and various organs. Good results have been shown in animal models and clinical trials...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/22A61L27/50A61L27/60
CPCA61L27/222A61L27/3633A61L27/50A61L27/60A61L2430/00C08L89/06
Inventor 杜亚楠李雅倩蒋书萌
Owner TSINGHUA UNIV
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