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Tissue-engineered atrioventricular node and construction method and application thereof

A technique of tissue engineering, construction method, applied in the field of biomedical engineering

Inactive Publication Date: 2014-04-23
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] At present, there are no research reports on the construction of tissue-engineered AV nodes at home and abroad, and there are only a few case reports on the use of engineered tissue constructs to establish AV conduction pathways

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1: Induction of canine bone marrow mesenchymal stem cells to prepare cardiac pacemaker cells and conduction cells

[0030] 1. Isolation and culture of canine bone marrow mesenchymal stem cells, canine sinus node cells and canine atrial appendage myocytes

[0031] (1) Isolation and culture of canine bone marrow mesenchymal stem cells

[0032]Bone marrow-derived mesenchymal stem cells were cultured by direct adherent method of whole canine bone marrow: extract 10 ml of iliac bone marrow from adult hybrid dogs (Experimental Animal Center of the Second Military Medical University, 15 kg), centrifuge at 900 × g for 10 min, discard the supernatant and fat; add red blood cells to lyse Liquid TrisNH 4 Cl, 4°C for 5 min to remove red blood cells, with 1 × 10 6 / ml was inoculated in fibronectin (FN) (Sigma company)-coated petri dishes, the medium was 60% DMEM-F12 (Gibco company), 10% FCS (Gibco company), 10ng / ml EGF (cytolab company) , 1000U / ml LIF (Chemicon Company). ...

Embodiment 2

[0043] Example 2: Induction of canine cardiac pacemaker cells and conduction cells seeded into collagen sponge to construct tissue engineered atrioventricular node

[0044] 1. Construction of tissue-engineered AV node

[0045] (1) Preparation of collagen sponge

[0046] Collagen was derived from type I collagen of bovine Achilles tendon, and the collagen sponge was provided by Wuxi Beidi Biological Co., Ltd. Cut the collagen sponge into 1.5×1.0×0.2cm size, use cobalt 60 Irradiate for more than 12 hours for disinfection.

[0047] (2) Collagen sponge inoculated with a mixture of canine cardiac pacemaker cells and conducting cells

[0048] Take the induced cardiac pacemaker cells and conduction cells respectively, add 1 ml of 0.25% trypsin (Gibco company), and keep the cells at 37°C and 5% CO. 2 Digest in an incubator for 1-2 minutes, observe under an inverted phase contrast microscope, the cell body retracts into a circle, add 1 ml of DMEM / F12 medium (Gibco) containing 10% F...

Embodiment 3

[0054] Example 3: Observation of conduction effect in vivo by tissue-engineered atrioventricular node transplantation

[0055] The successfully constructed tissue-engineered conduction bundles were transplanted into the subepicardium at the atrioventricular junction of dogs (Experimental Animal Center, Second Military Medical University, 15 kg) through thoracotomy, connecting the tissue-engineered conduction bundles to the coronary sulcus of the heart. In both atrioventricular myocardium, microsurgical suture; cyclosporine A (Beijing Shuanglu Pharmaceutical Co., Ltd., dose 0.25 mg / day) and prednisolone (Shanghai General Pharmaceutical Co., Ltd.) were started 3 days before transplantation Ltd., at a dose of 0.1 mg / day) to suppress immune rejection.

[0056] The results show that the tissue-engineered atrioventricular node of the present invention can survive in the host myocardium, and the cells in the transplanted tissue form an electromechanical coupling with the host myocard...

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Abstract

The invention relates to the technical field of biomedical engineering. At present, the research about the construction of a tissue-engineered atrioventricular node is not reported in China and foreign countries. The invention provides a method for constructing the tissue-engineered atrioventricular node by using cardiac pacing cells and conduction cells which are obtained by inducing collagen sponge and bone marrow mesenchymal stem cells. The bone marrow mesenchymal stem cells are induced into the pacing cells through co-culture with sinoatrial node cells, and are induced into the cardiac pacing cells by a method for co-culture with auricular appendage muscle cells; and the two kinds of cells are used as seed cells, the collagen sponge is taken as a bracket, and the tissue-engineered atrioventricular node with a pacing characteristic and an electrocardio-conduction characteristic is constructed in vitro. Through animal experiments, the successfully constructed tissue-engineered atrioventricular node is transplanted to treat atrioventricular conduction block, and a good treatment effect is achieved. The tissue-engineered atrioventricular node which has the electrocardio-conduction characteristic and can be transplanted for treatment is obtained, and hope is brought to treatment of arrhythmia caused by heart atrioventricular conduction block.

Description

technical field [0001] The invention relates to the technical field of biomedical engineering, in particular to a tissue-engineered atrioventricular node for treating heart atrioventricular conduction block disease and a preparation method thereof. Background technique [0002] The current idea of ​​biological pacing in the treatment of atrioventricular block is to use gene transfection or cell transplantation to establish a new stable pacemaker below the site of conduction block (such as the ventricular septum or the left and right ventricular walls) to provide and improve the ventricular Autonomic rhythm, or the introduction of pacemaker genes and pacemaker cells into the conduction block site to repair damaged conduction tissue, so that patients can avoid the implantation of electronic pacemakers and achieve the ideal treatment state advocated by biological cardiac pacing . However, although transfection of pacemaker genes improves and produces cardiomyocytes' self-disci...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/24
Inventor 李玉泉张传森张喜杨向群郭金萍张璐萍黄会龙刘镇姬瑞娟
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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