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Preparation method of precursor of composite tissues and organs with multichannel multilayer cell structure

A cell structure and composite tissue technology, applied in the direction of prosthesis, medical science, etc., can solve the problems of long time, difficult infiltration of cells, uneven distribution of cells, etc., and achieve the effect of excellent mechanical properties and excellent biocompatibility.

Inactive Publication Date: 2012-08-15
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The reconstruction of complex tissues and organs is realized by using the principles of mold combination and polymer coagulation forming. The invention can form complex three-dimensional structures containing different cell matrix materials and synthetic polymer scaffolds, which overcomes the need for inducing and culturing cells in three-dimensional scaffolds in tissue engineering. Long time, uneven cell distribution, it is difficult for cells to penetrate into the inner structure, etc.

Method used

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  • Preparation method of precursor of composite tissues and organs with multichannel multilayer cell structure
  • Preparation method of precursor of composite tissues and organs with multichannel multilayer cell structure
  • Preparation method of precursor of composite tissues and organs with multichannel multilayer cell structure

Examples

Experimental program
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Effect test

Embodiment 1

[0037] Embodiment 1: 1) Prepare a series of brass material matching combination molds with different sizes; 2) Prepare 50% (W / V) PLGA / 14dioxane solution, add 1% (W / W) Heparin is poured into the master mold. Use the dry film method of natural air drying to form the PLGA bottom support; 2) Prepare the fibrinogen solution, put the inner mold into the main mold, and inject the mixture of gelatin / fibrinogen and endothelial cells into the combined mold, and the cell density is 1× 10 7 3) Hepatocyte growth factor (HGF0.5ng / mL), human platelet-derived growth factor (BB or PDGF-BB 50ng / mL), transforming growth factor β1 (TGFβ1 10ng / mL) and basic fibroblast Cell growth factor (b-FGF 2.5ng / mL). Make the cell natural polymer material evenly distributed, inject thrombin solution (20IU / mL) to make the cell / natural polymer material layer form a stable structure; 4) remove the inner mold, and place the cell / natural polymer material layer on the surface and the inner mold The remaining gaps...

Embodiment 2

[0038] Example 2: 1) Silicon rubber is used to prepare a matching combined mold for multi-channel tissue and organ precursors; 2) 25% polyurethane / ethylene glycol solution is prepared, 5% paclitaxel is added, stirred evenly, and poured into the main mold. Use the cell culture fluid extraction method to form the outer polyurethane layer, and put it into the inner mold; 3) prepare the fibrinogen / endothelial cell mixture containing 5% paclitaxel (the cell density is 1 × 10 7 pcs / mL), perfuse into the combination mold and add thrombin solution (20IU / mL) to form the first layer of main material; 4) inject the mixture of gelatin / fibrinogen and adipose stem cells on the first layer of main material, the cell density 1×10 6 cells / mL to form the second layer of main material; 5) inject fibrinogen / hepatocyte mixture (cell density is 1×10) on the second layer of main material 7 pcs / mL), soak the molded object with thrombin solution (10IU / mL) for 1 minute to form a three-layer structure ...

Embodiment 3

[0039] Embodiment 3: 1) Prepare a series of polytetrafluoroethylene matching combination molds with different sizes; 2) Prepare a polylactic acid / isopropanol solution with a concentration of 30%, add 30% sodium citrate, stir evenly, pour into inside the master mold. Use the PBS extraction method to form the bottom support, and put it into the inner mold; 3) prepare the collagen / endothelial cell mixture of 1% sodium citrate (the cell density is 1×10 7 per mL), poured into the combination mold, and placed at 37°C for 10 minutes to stabilize the structure of the collagen / endothelial cell mixture and form the first layer of main material; 4) inject the collagen / smooth muscle cell mixture on the first layer of main material ( Cell density is 1 x 10 7 cells / mL); placed at 37°C for 10 minutes to stabilize the structure of the collagen / smooth muscle cell mixture and form the second layer of main material; 1), the cell density is 1×10 6 cells / mL, placed in an incubator at 37°C for 1...

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Abstract

The invention discloses a preparation method of a precursor of composite tissues and organs with a multichannel multilayer cell structure. The preparation method comprises the following steps that: one or more cell matrix solution and synthetic macromolecule solution are prepared, the synthetic macromolecule solution is injected into a main mold to form the lower surface of a support, and then the one or more cell matrix solution is injected into an assembling mold in a layered manner to form a plurality of cell matrix layers through crosslinking; and an inner mold is removed, the synthetic macromolecule solution is injected into the outer surface and the surrounding gap of the cell matrix layers to form an outer support, and the assembling mold is removed. The preparation method can be used for forming a complex multichannel three-dimensional structure combining different cell matrix materials containing uniform internal channels with a case of the synthetic macromolecule support, and overcomes the defects of tissue engineering that when cells are induced and cultured in the three-dimensional support, the needed time is long, the type of the cells is single, the distribution of the cells is nonuniform, the cells closed in a cell layer which exceeds a certain thickness are not easy to survive, nutrient channels do not exist, the layering differentiation on the stem cells is difficult to realize, the limitation on the appearance of the support is strong, and the like.

Description

technical field [0001] The invention belongs to the technical field of artificial manufacturing of biological tissues and organs, in particular to a process for preparing tissue and organ precursors by using synthetic polymer materials and cell matrix materials, and belongs to the technical field of biological tissue engineering. Background technique [0002] Tens of millions of patients suffer from tissue defect or organ failure every year in the world. However, living donor organs are limited, and existing mechanical devices do not have all the functions of the organs and cannot prevent further deterioration of the patient's condition. Accordingly, the tissue engineering (Tissue Engineering) technology for the purpose of improving the treatment level of such diseases came into being. [0003] Tissue engineering was officially proposed and determined by the National Science Foundation of the United States in 1987. It is the application of the principles of cell biology, bi...

Claims

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Application Information

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IPC IPC(8): A61L27/44A61L27/38
Inventor 王小红
Owner TSINGHUA UNIV
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