Method for constructing tissue-engineered pulsating tissue

A technology of pulsating tissue and tissue engineering, applied in the field of biomedical engineering technology and regenerative medicine

Inactive Publication Date: 2014-12-24
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] At present, there is no research report on the construction of tissue engineered beating tissue at home and abroad, only Choi et al. Using skeletal muscle cells and matrigel to construct an organoid in vitro, and transplanting it into the subepicardium of the coronary sulcus of the rat heart, a preliminary attempt was made to rebuild the atrioventricular conduction pathway

Method used

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  • Method for constructing tissue-engineered pulsating tissue
  • Method for constructing tissue-engineered pulsating tissue

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1: Inducing mouse brown adipose tissue-derived adipose-derived stem cells into pacemaker cells

[0049] 1. Isolation and culture of adipose-derived stem cells

[0050] (1) Cut the skin between the scapula of the mouse, remove the white fat layer covered by the brown adipose tissue between the scapula, take about 0.4 g of brown fat, wash it in PBS at 4°C for 3 times, transfer it to a 5ml centrifuge tube, and add 0.8ml0 .1% type II collagenase in high-sugar DMEM (containing 1% BSA), shredded.

[0051] (2) Transfer the shredded adipose tissue to another centrifuge tube, add 10ml of 0.1% type II collagenase high-sugar DMEM (containing 1% BSA), digest at 37°C for 45min, and shake and mix repeatedly during this time. Centrifuge at 3000rpm for 1min, suck the supernatant through a 100-mesh filter, and enter (4).

[0052] (3) Add 5ml of 0.1% type II collagenase high-sugar DMEM (containing 1% BSA) to the above centrifuge tube, digest at 37°C for 20min, and shake and mix...

Embodiment 2

[0064] Example 2: Implantation of induced pacemaker cells into collagen sponges to construct tissue-engineered beating tissues

[0065] 1. Construction of pacemaker cell-collagen sponge complex

[0066] (1) Preparation of collagen sponge: Collagen sponge was directly purchased from a commercial company (Wuxi Beidi Biological Co., Ltd.), the collagen sponge was cut into a size of 0.8×0.6×0.2cm, 60 Irradiate for more than 12 hours for disinfection.

[0067] (2) Preparation of pacemaker cells: 0.25% trypsin digested adipose-derived stem cells (more than 90% had been differentiated into pacemaker cells), 5min, centrifuged at 1500rpm for 5min. Discard the supernatant, make a cell suspension, and adjust to a cell density of 1.0×10 7 pieces / ml.

[0068] (3) Construction of pacemaker cell-collagen sponge complex: under a stereomicroscope, use a micro-syringe to evenly inoculate the cell suspension on the collagen sponge at 4 points, the total amount is about 100 μl, and the cell d...

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Abstract

The invention relates to the fields of biomedical engineering technology and regenerative medicine and particularly relates to a method for constructing a tissue-engineered pulsating tissue. According to the method disclosed by the invention, stem cells derived from brown adipose are induced into sinuatrial-node-like cells, and the sinuatrial-node-like cells are implanted onto collagen sponge to construct the tissue-engineered sinuatrial-node-like pulsating tissue in vitro. The tissue-engineered tissue with pulsating characteristics obtained by virtue of the method provides a tissue model for a research on screening of arrhythmic drugs and brings hope for treatment of arrhythmic diseases caused by sick sinus syndrome and auriculo-ventricular block.

Description

technical field [0001] The invention relates to the fields of biomedical engineering technology and regenerative medicine, in particular to a method for preparing tissue-engineered beating tissue. Background technique [0002] At present, the methods for constructing cardiac biological pacemaker mainly include ① using gene transfection to modulate the ionic current of cardiomyocytes; ② implanting pacemaker cells or engineered pacemaker cells into the heart. [0003] Among these methods, the method of gene transfection has the possibility of causing cardiac arrhythmia. Moreover, exogenous genes are difficult to effectively regulate in vivo, and viral vectors remain in the body. Cell transplantation can overcome the defects of gene transfection, but the transplanted cells are very easy to migrate and diffuse in the heart, and cannot form a stable and uniform cardiac rhythm point, and the cells currently used for transplantation are not suitable for clinical biological cardiac...

Claims

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Application Information

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IPC IPC(8): C12N5/0775A61F2/02A61L27/38A61K49/00
Inventor 杨向群陈磊姬瑞娟李玉泉刘芳邓子军张喜张传森
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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