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Acellular matrix and preparation method thereof

A technology of acellular matrix and cells, which is applied in the field of acellular matrix and its preparation, can solve the problems of cell destruction, loss of extracellular matrix components, and destruction of extracellular matrix ultrastructure, and achieve the effect of convenient removal

Active Publication Date: 2015-03-11
欧亚科(广东)生命科学有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The physical method refers to the destruction of cells by physical methods such as freezing, high pressure, ultrasound, osmotic pressure change, and electric current. At the same time, it also destroys the ultrastructure of the extracellular matrix to varying degrees, and usually needs to be combined with other decellularization methods
The reason is that in theory, the existing decellularization schemes inevitably lead to the loss of extracellular matrix components, especially soluble proteoglycan and glycoprotein components, and the loss rate can reach 80%.

Method used

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Examples

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preparation example Construction

[0032] The method for preparing the acellular matrix of the present invention mainly comprises:

[0033] a. Pretreatment of tissues and organs to be used:

[0034] At room temperature, according to the principle of conventional aseptic operation, the tissues and organs to be used are taken out, and the pretreatment methods can be different according to the different tissues and organs to be taken. Basically include: routine cleaning, disinfection, and separation of tissues and organs to be used with a physiological buffer solution containing antibiotics. The purpose of this step is to remove microorganisms and meet the aseptic principle during subsequent tissue culture. At the same time, the pretreatment process needs to ensure a high cell survival state in fresh tissues (viable cells>99%).

[0035] b. In the culture environment, apply apoptosis-inducing factors to the tissues and organs to be used to promote the spontaneous formation of apoptotic bodies in living cells;

[...

Embodiment 1

[0042] Example 1 Preparation of decellularized pig mesentery matrix

[0043] 1. Product preparation

[0044] The following processes are all aseptic operations, and the reagents used are all sterilized. The method for preparing decellularized porcine mesentery matrix described in this embodiment comprises the following steps:

[0045] a. Pretreatment of tissues and organs to be used;

[0046] Under normal aseptic operation at room temperature, fresh porcine mesentery tissue (5 cm x 5 cm) was removed. Soak the obtained porcine mesentery tissue in carbonate buffer solution containing antibiotics (100 U / ml penicillin G, 100 μg / ml streptomycin sulfate) for 3 to 5 times, each time for 5 to 10 minutes.

[0047] b. Apply the inducing factors that promote cell apoptosis to the tissues and organs to be used, place them in the culture environment, and promote the spontaneous apoptosis of living cells to form apoptotic bodies; (cytokine induction)

[0048] Place fresh porcine mesente...

Embodiment 2

[0072] Example 2 Preparation of Decellularized Porcine Pericardium Matrix

[0073] 1. Product preparation

[0074] The following processes are all aseptic operations, and the reagents used are all sterilized. The method for preparing decellularized porcine pericardium matrix described in this embodiment comprises the following steps:

[0075] Prepare experimental group

[0076] a. Pretreatment of tissues and organs to be used;

[0077] Under normal aseptic operation at room temperature, fresh porcine pericardium tissue (5 cm x 5 cm) was removed. Soak the obtained porcine pericardium tissue in carbonate buffer solution containing antibiotics (100 U / ml penicillin G, 100 μg / ml streptomycin sulfate) for 3 to 5 times, each time for 5 to 10 minutes.

[0078] b. Apply the inducing factors that promote cell apoptosis to the tissues and organs to be used, place them in the culture environment, and promote the spontaneous apoptosis of living cells to form apoptotic bodies; (ray + gr...

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Abstract

The invention provides a new acellular matrix and a preparation method thereof. The preparation method comprises the following steps: preprocessing a tissue organ to be used, wherein the tissue organ to be used is a heterogeneous, or homogeneous xenogenous or autologous tissue organ; in culturing environment, exerting apoptosis induction factors on the tissue organ to be used so as to promote living cells to spontaneously form apoptotic bodies; eliminating the apoptotic bodies in the tissue organ to be used; cleaning the prepared acellular matrix. The method disclosed by the invention adopts a driving apoptotic cell removing manner, so that the destructive side effects of too much physical and chemical factors exerted by outside on extracellular matrix components are reduced, the spontaneous separation and degradation of cell components can also be effectively realized, and great convenience is brought for the elimination of subsequent cell components.

Description

technical field [0001] The invention belongs to the field of tissue engineering, and in particular relates to an acellular matrix and a preparation method thereof. Background technique [0002] Various bioscaffold materials derived from acellular matrices have been widely used in tissue engineering and regenerative medicine research, and have shown potential for clinical application in human diseases. By removing heterogeneous or allogeneic cells in various tissues and organs, and retaining the complex components and precise structures, the acellular matrix of the corresponding tissues and organs can be obtained. In theory, any decellularization method will affect the composition and structure of the natural tissue matrix to varying degrees, and ultimately affect the host's response to the acellular matrix after transplantation. How to optimize the decellularization method and improve the decellularization process is still an important part of the research in this field. ...

Claims

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Application Information

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IPC IPC(8): A61L27/36
Inventor 武征张建华
Owner 欧亚科(广东)生命科学有限公司
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