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Tissue engineering myocardium bioreactor constructed by pouring, perfusion and pulsation combination

A technology of bioreactor and tissue engineering, applied in the direction of specific-purpose bioreactor/fermenter, biochemical instrument, biochemical equipment and method, etc., can solve the problem of ambiguous relationship between mechanism design, failure to maintain and repeated use, Cardiomyocytes are difficult to establish connections, etc., to achieve the effect of increasing cell planting density, strengthening exchange, and promoting directional connections

Inactive Publication Date: 2012-05-02
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] (3) The tissue-engineered myocardium lacks an environment similar to the real body in the in vitro culture process, and it is difficult for cardiomyocytes to establish connections and spread to form tissues
The possible problem of perfusion bioreactor in myocardial tissue culture is: the shear stress caused by the liquid flow to the cells may cause certain damage to the cells
[0012] In the existing reports and invention patents, there are reports on cell perfusion, perfusion and pulsation related bioreactors, but due to the relatively large contradictions and conflicts in the design of their mutual mechanisms, and the unclear interaction relationship, etc. There is no bioreactor that can achieve multi-factor joint culture. The main reason is that the environment and mechanism of cell perfusion, perfusion culture and stress training are different. In terms of the structure of the bioreactor, the structural design problem cannot be well solved to ensure their respective effects. The effects are all achieved normally, and it is easy to cause problems such as complex structure of the reactor, inability to maintain and reuse, and inability to match the operating parameters of various functions.

Method used

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  • Tissue engineering myocardium bioreactor constructed by pouring, perfusion and pulsation combination
  • Tissue engineering myocardium bioreactor constructed by pouring, perfusion and pulsation combination
  • Tissue engineering myocardium bioreactor constructed by pouring, perfusion and pulsation combination

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0053] Example 1: Construction of tubular tissue-engineered myocardium

[0054] Liquid collagen (1%) is mixed with chitosan (1% acetic acid solution) to obtain a tissue engineering scaffold material, and a pipe-shaped mold is used to solidify at a low temperature and freeze-dried for 24 hours to obtain a pipe-shaped scaffold. The scaffold was cross-linked with sodium polyphosphate and adjusted to be neutral, rinsed with PBS and dried for later use. Install the bracket on the pulsation effect hose, and then install and shape the bioreactor as a whole. The newly isolated mouse cardiomyocytes were dispersed in the culture medium to obtain a cell suspension, and the concentration of the suspension was 10 6 cells / mL. At 4°C, 1ml of the cell suspension was mixed with 1ml of 10% Metrigel solution, and the mixed solution was added into the perfusion chamber, and after the liquid inlet was closed, the cells were perfused for 20 minutes. Cells were cultured statically after perfusion...

Embodiment 2

[0055] Example 2: Construction of Strip-shaped Tissue-Engineered Myocardium Patch

[0056] Mix according to the method in Example 1 to obtain collagen-chitosan tissue engineering scaffold material, use a strip-shaped mold, freeze-dry for 24 hours after solidifying at a low temperature to obtain a strip-shaped scaffold. The scaffold was cross-linked with sodium polyphosphate and adjusted to be neutral, rinsed with PBS and dried for later use. Wrap the support around the pulsation effect hose along the length direction, and then install and shape the bioreactor as a whole. The newly isolated mouse cardiomyocytes were dispersed in the culture medium to obtain a cell suspension, and the concentration of the suspension was 10 6 cells / mL. At 4°C, 1ml of the cell suspension was mixed with 1ml of 10% Metrigel solution, and the mixed solution was added into the perfusion chamber, and after the liquid inlet was closed, the cells were perfused for 20 minutes. Cells were cultured stati...

Embodiment 3

[0057] Example 3: Effect of cell perfusion on the construction of tissue engineered myocardial patch

[0058] Mix according to the method in Example 1 to obtain collagen-chitosan tissue engineering scaffold material, and prepare a pipeline-shaped scaffold. The scaffold was cross-linked with sodium polyphosphate and adjusted to be neutral, rinsed with PBS and dried for later use. Install the bracket on the pulsation effect hose, and then install and shape the bioreactor as a whole. The newly isolated mouse cardiomyocytes were dispersed in the culture medium to obtain a cell suspension, and the concentration of the suspension was 10 6 cells / mL. At 4°C, 1ml of the cell suspension was mixed with 1ml of 10% Metrigel solution, and the mixed solution was added into the perfusion chamber, and after the liquid inlet was closed, the cells were perfused for 20 minutes. Then carry out static culture, and replace the culture medium every 24 hours. After culturing for 3 days, it can be ...

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Abstract

The invention relates to a tissue engineering myocardium bioreactor constructed by pouring, perfusion and pulsation combination, belonging to the technical field of tissue engineering and biological engineering manufacture. The bioreactor comprises a cell pouring generator, a perfusion module, a pulsation generator, a culture module and a control module; the cell pouring generator, the perfusion module and the pulsation generator are communicated with the culture module through pipelines; and the culture module comprises a pulsation cavity, a pulsation effect soft tube with one closed end anda perfusion cavity, and the perfusion module and the pulsation generator carry out perfusion culture and pulsation training. The invention can improve cardiomyocytes planting depth and simulate myocardium in vivo environment, and carries out culture to obtain a tissue engineering myocardium with larger thickness and autonomously beating capacity. The bioreactor is used for realizing that the tissue engineering myocardium construction carries out perfusion culture and pulsation training simultaneously or separately on cell perfusion and tissue culture stages in a cell planting stage.

Description

technical field [0001] The invention belongs to the field of tissue engineering and biomanufacturing, and relates to a tissue engineering myocardial bioreactor constructed by perfusion-perfusion-pulsation, which simultaneously realizes the cell perfusion in the cell planting stage and the perfusion culture in the tissue culture stage in the in vitro culture process of tissue engineered myocardium And pulsation training, simulating the internal environment, its main operating parameters can be automatically adjusted and controlled. Background technique [0002] The heart is the power organ of the human blood circulation and an important organ to maintain the life activities of the human body. At present, various heart diseases have become the most terrible killer of human health. Many current clinical treatments have certain limitations. Myocardial tissue engineering is a research hotspot in the field of tissue engineering. Its basic principle is to construct a three-dimens...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/00
CPCC12M29/10C12M35/04C12M21/08
Inventor 熊卓胡京奇张婷张人佶林峰颜永年
Owner TSINGHUA UNIV
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