Prepn process of complicated tissue organ precursor

A technology for complex tissues and organs, applied in the field of preparation of complex tissue and organ precursors, can solve problems such as aging, failure to treat clinical patients in time, and failure to meet accurate positioning and fixed-point placement

Inactive Publication Date: 2005-04-27
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing tissue engineering technology faces many difficulties and limitations. The success of tissue engineering application research is in those tissues and organs with relatively simple structure and physiological functions, such as bone, cartilage, skin, etc.
The traditional stent preparation technology cannot accurately control the size, structure, spatial distribution and through channels of the pores, and the supply of nutrients and the growth of blood vessels are greatly restricted.
The traditional tissue engineering method generally prepares the structural scaffold first. During the cell culture process, the cells in the upper layer consume most of the oxygen and nutrients, which limits the diffusion of these components to the bottom layer, thereby limiting the migration of cells to the deep layer of the scaffold, etc.
This method of preparing the scaffold first and then culturing the cells is time-consuming and laborious. The cells are likely to be deformed and aged during the process of migrating into the scaffold, which cannot meet the requirements of timely treatment of clinical patients.
At the same time, traditional tissue engineering technology cannot meet the needs of accurately positioning and placing different cells in space and constructing functionally graded structures of complex tissues and organs.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Example 1: Preparation of artificial composite liver precursor

[0021] 1) Purchasing existing cell lines such as HepG2 hepatocytes and endothelial cells, and centrifuging the two types of cell suspensions respectively with 1% gelatin / sodium alginate (100:1w / w) 0.09M NaCI solution and cell growth factors well mixed. Spray into a 5% calcium lactate solution by weight with an electrostatic microcapsule generator to form microspheres. Aspirate the calcium lactate solution, add 0.1% glutaraldehyde solution to react for 1 second to form microgel beads; react the microgel beads with 0.1% by weight sodium citrate (also called sodium citrate) buffer solution for several minutes , with liquefied microbead cores to form liquefied microcapsules.

[0022] 2) Preparation of network-like structures: the biodegradable polymer material polyurethane (PU) 1,4-dioxane solution with a biological concentration of 1% is used to make three-dimensional images of different parts of network-sh...

Embodiment 2

[0024] Example 2: Preparation of artificial composite liver precursor

[0025] 1) Purchasing existing cell lines such as HepG2 liver cells and endothelial cells. After the two cell suspensions were centrifuged, they were mixed evenly with 20% gelatin / sodium alginate (1:1 w / w) 0.09M NaCI solution and endothelial cell growth factor respectively.

[0026] 2) Preparation of network-like structures: the biodegradable polymer material polyurethane (PU) 1,4-dioxane solution with a biological concentration of 1% is used to make three-dimensional images of different parts of network-shaped tissues and organs using computer-aided rapid prototyping technology. structural support;

[0027] 3) Assembling: the two kinds of cell-matrix complexes prepared in step 1) were injected into the corresponding positions of the three-dimensional structure scaffold by micro-droplet injection technology, and cross-linked with 5% calcium lactate solution by weight. Aspirate the calcium lactate solution...

Embodiment 3

[0028] Example 3: Preparation of artificial composite liver precursor

[0029] Isolate the patient's liver cells, endothelial cells and stellate cells, culture and expand them in vitro. The three cell suspensions were centrifuged separately or mixed with 0.09M NaCI solution of 10% gelatin / chitosan (1:100w / w), hepatocyte growth factor and endothelial cell growth factor, prepared by cell assembly machine from endothelial cells And the multi-channel three-dimensional structure composed of hepatocytes and stellate cell layers. React with 1% glutaraldehyde solution for 1 second, and rinse with normal saline. This structure can be directly implanted in the intestinal membrane or on the site of liver injury, and can also be implanted after compounding with the tubular network polyurethane with endothelial cells inside.

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Abstract

The present invention is preparation process of complicated tissue organ precursor and belongs to the field of biological tissue engineering technology. Seed cell or gene and matrix material are prepared into micro colloid bead or 3D multi-canal structure; and cell and rack are located in space accurately via fast computer aided formation, coating, injecting, etc. The said process is superior to available tissue engineering process, which is time consuming and has uneven distribution of cells in rack, hard permeation of cell to deep structure and other demerits. By means of part combination, the present invention can reach the requirement of complicated organ, and realize organ reconstruction for repair and regeneration.

Description

technical field [0001] The invention relates to a method for preparing complex tissue and organ precursors, belonging to the technical field of biological tissue engineering. Background technique [0002] There are tens of millions of patients suffering from tissue defect or organ failure every year in the world, and about 8 million of these patients are treated with surgery every year in the United States alone. 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, Tissue Engineering (Tissue Engineering) technology with the purpose of improving the treatment level of such diseases came into being. Tissue engineering is a high-tech discipline produced by the intersection of biology, medicine, materials science, engineering and other disciplines. Its meaning is to apply the principles and technologies of life science and engineeri...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071
Inventor 颜永年王小红熊卓林峰吴任东张人佶卢清萍
Owner TSINGHUA UNIV
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