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Method and apparatus for constructing blood vessel in vitro in the tissue project

A technology of tissue engineering and construction method, which is applied in the direction of tissue cell/virus culture device, tissue culture, biochemical cleaning device, etc. It can solve the problems of inability to achieve the structural strength of blood vessels, lack of elastic medial support of blood vessels, and shedding of endothelial cell layer, etc. problem, to achieve the effect of simple structure, promotion of directional arrangement, and increase of shrinkage characteristics

Inactive Publication Date: 2006-01-11
上海组织工程研究与开发中心
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The technical problem to be solved in the present invention is to disclose a method for constructing tissue engineered blood vessels in vitro and a device for constructing blood vessels in vitro, so as to overcome the lack of sufficient elastic medial support in blood vessels in the prior art, which cannot reach or approach the structural strength of normal blood vessels, and The implanted endothelial cell layer cannot withstand the erosion of blood flow and falls off, causing secondary thrombosis and reducing its long-term patency rate and other defects

Method used

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  • Method and apparatus for constructing blood vessel in vitro in the tissue project
  • Method and apparatus for constructing blood vessel in vitro in the tissue project
  • Method and apparatus for constructing blood vessel in vitro in the tissue project

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Embodiment 1

[0041] (1) Cell culture and expansion:

[0042]Endothelial cells: samples were obtained using the enzymatic digestion method disclosed in the document "Jones GE. Methods in Molecular Medicine Human cell Culture Protocols." Perfuse the human umbilical vein or animal vein with 0.1wt% collagenase, digest at 37°C for 15 minutes, collect the digestion solution, inoculate it in a multi-well plate, and place it in 5% CO 2 Cultivate in an incubator. After the colony is formed, change the medium once every 2 to 3 days;

[0043] Smooth muscle cells: the tissue patch method disclosed in the above literature was used. After carefully separating the obtained arteries, the middle layer was obtained and cut into a size of about 1-2mm 2 Paste the small pieces in the Petri dish. After the paste is firm, add the culture medium and place it in a concentration of 5% CO 2 Cultivate in an incubator and change the medium every 2-3 days.

[0044] (2) Identification of cell properties:

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Abstract

The present invention discloses the method and apparatus for externally constructing blood vessel tissue engineering. Biomechanical factors are applied to stimulate proliferation of blood vessel smooth muscle, promote directional arranging of blood vessel, increase the contraction characteristic of smooth muscle cell, increase the collagen content in cell-material composition and the expression of actin heavy strand in cell and induce endothelial cell to form stress fiber via stress. The apparatus of the present invention includes reactor and controller and the controller has its signal port connected via wire to solenoid valve to form pulsed water flow and flow rate to simulate internal physiological environment. The apparatus has reasonable structure, stable performance and easy operation, and can perform data processing and analysis can provide enclosed, non-toxic, bacteria-free and oxygen-rich environment for external blood vessel construction.

Description

technical field [0001] The invention relates to an in vitro construction method of tissue engineered blood vessels and a construction device adopted, in particular to an in vitro construction method and a bioreactor capable of simulating the pulsation stimulation of human blood vessels and the flushing effect of blood flow. Background technique [0002] With the increase in the incidence of hypertension, diabetes, etc., the incidence of peripheral vascular disease is increasing year by year. There are more than 1 million patients who need vascular transplantation due to vascular disease, and 60% of them require small blood vessels with a diameter of less than 6mm. People try Autologous arteries and veins, allografts, xenograft arteries and veins, artificial blood vessel endothelialization and other methods. However, these methods have more or less deficiencies in terms of long-term effects. The concept of tissue engineering in the late 1980s and its rapid development inject...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/08C12M3/00C12M1/00A61L27/00A61L27/28A61L27/14
Inventor 曹谊林李宏刘阳刘伟崔磊
Owner 上海组织工程研究与开发中心
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