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Method of external constructing tissue engineering blood vessel

A technology of tissue engineering and blood vessels, applied in the field of tissue engineering and materials science, can solve the problem of flat and less elastic lumen

Active Publication Date: 2005-08-10
上海组织工程研究与开发中心
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Niklason (Niklason LE, Gao J, Abbott WM, et al.Functional arteries grown in vitro.Science 1999; 284:489-496) and others have reported using vascular bioreactors, using PGA-cell complexes, and culturing in vitro for 6-8 weeks , to construct tissue-engineered blood vessels, but it adopts the hemodynamic conditions of mammalian embryonic development to cultivate blood vessel wall cells from adult mammals, so the lumen is generally flat and less elastic, and its histological structure is similar to that of normal blood vessel tissue. there is still a certain gap

Method used

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  • Method of external constructing tissue engineering blood vessel
  • Method of external constructing tissue engineering blood vessel
  • Method of external constructing tissue engineering blood vessel

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preparation example Construction

[0051] The preparation method of the tissue-engineered blood vessel of the present invention is simple. A certain number of smooth muscle cells are inoculated on a pharmaceutically acceptable biodegradable material, and then the cell-material compound is wrapped in an elastic material tube; Conditionally cultivate the cell material complex of step (b) for 3-10 days (more preferably 5-8 days); then apply the following dynamic stimulation through the elastic material tube, and continue to cultivate for 5-50 days (preferably 7-30 days) day): beat frequency: 75±10 beats / min; dilatation: 5±1%; pressure: 0.001-0.04Mpa; flow: 0.01-100ml / min, thereby obtaining tissue-engineered vascular grafts.

[0052] The shape of the tissue engineered vascular graft of the present invention is usually ring. The thickness of the vascular graft of the present invention is not particularly limited, and is usually 5-200 microns, preferably 10-50 microns, more preferably 20-40 microns.

[0053] The smo...

Embodiment 1

[0063] Isolation, culture, expansion and observation of vascular smooth muscle cells

[0064] Select young Changfeng hybrid pigs, male or female, weighing 10-15kg, aged 2 months. Under a sterile environment, a section of porcine common carotid artery was surgically excised, 3-4cm long, after peeling off the adventitia and scraping off the intima, the arterial media was cut into 1×1×1mm 3 Spread the tissue pieces evenly on the culture dish, add DMEM culture solution containing 10% fetal bovine serum (FBS) after adhering to the wall, place at 37°C, 5% CO 2 The cells were cultured in an incubator, and the second generation cells were used for experiments. Cell growth was observed with an inverted phase-contrast microscope, and samples were taken for immunohistochemical detection of the expression of α-smooth muscle actin, and its ultrastructure was observed with a transmission electron microscope.

[0065] result:

[0066] (a) Morphological observation of cells

[0067] On th...

Embodiment 2

[0073] Smooth muscle cell identification

[0074] Immunohistochemical staining for detection of α-smooth muscle actin: smooth muscle cell slides were divided into experimental group and blank control group, and human skin fibroblast slides were retained as negative control group. Main steps Tissue sections were fixed in anhydrous acetone, treated with 0.25% TritonX-100 to increase cell permeability, treated with 1.5% H2O2 to block endogenous peroxidase, incubated with sheep serum to remove non-specific staining, and dropped primary antibody (mouse Anti-human smooth muscle α-actin antibody diluted 1:50 (DAKO company, Denmark), PBS was added dropwise to the blank control group, 4 ° C refrigerator overnight, and the secondary antibody (horseradish peroxidase-labeled goat anti-mouse IgG) was added dropwise. React at 37°C for 30 minutes, develop color with DAB, stain with hematoxylin, dehydrate with gradient alcohol, and seal with neutral resin. Judgment of the results: the cells ...

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Abstract

A process for in vitro configuration of histoengineered blood vessel and its smooth muscle are disclosed. The resultant histoengineered blood vessel has better elasticity, bright lustre and dense elastic fibres.

Description

technical field [0001] The invention relates to the fields of tissue engineering and material science, and more particularly relates to a method for constructing tissue engineered blood vessels in vitro and the constructed tissue engineered blood vessels. Background technique [0002] Seeking an ideal vascular graft is a major clinical issue that needs to be solved urgently. The rise of tissue engineering research has brought dawn to the solution of this difficult problem. [0003] In previous studies, Weinberg, Shinoka, L'heureux, and Bader used different methods to build tissue-engineered blood vessels in vitro, and tried to use them to repair animal vascular defects (Weinberg CB and Bell E.A blood vessel model constructed from collagen and cultured vascular cells. Science 1986; 23: 397-340; Shinoka T, Tim DS, Ma PX, et al. Creation of viable pulmonary artery autograftsthrough tissue engineering. J Thorac Cardiovasc Surg 1998; 115: 536-545; Bader A, Steinhoff G, Strobl K,...

Claims

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Application Information

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IPC IPC(8): A61F2/06C12N5/08C12N11/02C12N11/08
Inventor 曹谊林崔磊刘伟许志成李宏刘阳
Owner 上海组织工程研究与开发中心
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