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Extracellular matrix freeze-drying powder

An extramatrix, freeze-dried powder technology, applied in the field of cell biology, to achieve the effect of preventing changes, low moisture content, and improving stability

Inactive Publication Date: 2018-11-16
广州昕生医学材料有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

There are currently no related extracellular matrix products on the market

Method used

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  • Extracellular matrix freeze-drying powder

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0036] An embodiment of the freeze-dried extracellular matrix powder provided by the present invention, the preparation method of the freeze-dried extracellular matrix powder comprises the following steps:

[0037] (1) Take the porcine small intestine tissue, wash after defatting and decellularization, and obtain the crude extracellular matrix extract;

[0038] (2) Mix the acid protease with a concentration of 0.01% and the crude extract of the extracellular matrix at a weight ratio of 1:20, shake for 36 hours, centrifuge to take the supernatant, use a dialysis bag with a molecular weight cut-off of 50,000Da, and dialyze with running water for 24 hours , until the conductivity is 1.5um / s, the extracellular matrix extract is obtained;

[0039] (3) Filter the extracellular matrix extract obtained in step (3) using a 0.8 μm microporous membrane, mix the filtrate with mannitol, and make the concentration of mannitol in the mixed solution 10%, and the mixed solution is heated at -2...

Embodiment 2

[0041] An embodiment of the freeze-dried extracellular matrix powder provided by the present invention, the preparation method of the freeze-dried extracellular matrix powder comprises the following steps:

[0042] (1) Take porcine pericardium tissue, wash after degreasing and decellularization to obtain crude extracellular matrix extract;

[0043] (2) Mix pepsin with a concentration of 1% and the crude extracellular matrix extract according to a weight ratio of 1:5, shake for 12 hours, filter the filtrate, add ethanol to the filtrate to make the final concentration 60%, Stir evenly, and obtain the extracellular matrix extract after standing still for precipitation;

[0044] (3) Centrifuge the extracellular matrix extract obtained in step (3), mix the supernatant with glucose so that the concentration of glucose in the mixed solution is 0.2%, and freeze the mixed solution at -20°C for 3 hours, and then freeze it at - Freeze-dry at 80° C. for 12 hours to obtain the freeze-drie...

Embodiment 3

[0046] An embodiment of the freeze-dried extracellular matrix powder provided by the present invention, the preparation method of the freeze-dried extracellular matrix powder comprises the following steps:

[0047] (1) Take the bovine bladder tissue, degrease and wash after decellularization to obtain the crude extracellular matrix extract;

[0048] (2) Mix the neutral protease with a concentration of 0.1% and the crude extract of the extracellular matrix according to a weight ratio of 1:10, after shaking for 24 hours, centrifuge to get the supernatant, add sodium chloride to the supernatant to make it The final concentration is 0.5mol / L, stir evenly, and obtain the extracellular matrix extract after standing for precipitation;

[0049] (3) Filter the extracellular matrix extract obtained in step (3) with a 0.8 μm microporous membrane, mix the filtrate with trehalose, and make the concentration of trehalose in the mixed solution 1%, and the mixed solution is heated at -20°C F...

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Abstract

The invention discloses an extracellular matrix freeze-drying powder. The extracellular matrix freeze-drying powder is prepared by a method with the following steps of (1) selecting animal tissues, degreasing, removing cells, and cleaning, so as to obtain a crude extract of the extracellular matrix; (2) extracting the crude extract of the extracellular matrix to obtain a hydrolysate, centrifugingor filtering the hydrolysate, and treating a supernatant or filtrate by a dialyzing method, an organic settling method or a salt settling method, so as to obtain the extract of the extracellular matrix; (3) filtering or centrifuging the extract of the extracellular matrix, mixing the filtrate or supernatant and monosaccharide or disaccharide, and freeze-drying, so as to obtain the extracellular matrix freeze-drying powder. The extracellular matrix freeze-drying powder has the advantages that the multiple active matters of polytype collagen, proteoglycan, growth factors and the like in the animal tissues are reserved; the multiple biological activities are realized, the biocompatibility is good, and the tissue regeneration is promoted; the anti-oxidizing, repair and wrinkle-resisting effects are realized, and the proliferation, transfer and differentiation of the cells are induced; the extracellular matrix freeze-drying powder can be used as the structure support, and the filling function is realized.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and in particular relates to a freeze-dried extracellular matrix powder. Background technique [0002] The extracellular matrix is ​​a macromolecule synthesized and secreted by animal cells, distributed on the cell surface or between cells, and can be divided into three categories: (1) Glycosaminoglycans and proteoglycans, which can form water-based (2) Structural proteins, such as collagen and elastin, which endow the extracellular matrix with certain strength and toughness; (3) Adhesive proteins, also known as fibronectins, such as fibronectin and laminin, which can Facilitates the binding of cells to the matrix. [0003] Amino polysaccharides can be divided into six types according to the composition of sugar groups, connection methods, sulfation degree and position, namely hyaluronic acid, chondroitin sulfate, dermatan sulfate, heparan sulfate, heparin and keratan sulfate. Among them, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0226
Inventor 杨伟红耿梦梦姜玉峰程飚苏玉天刘江辉陈妮丹陈大鹏邓小敏
Owner 广州昕生医学材料有限公司
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