Method for extracting matrix gel from cattle and donkey skin

A skin and gel technology, applied in the field of extracting matrix gel, can solve the problems of single component and single biological function, and achieve the effect of promoting tissue repair

Pending Publication Date: 2021-12-21
HENAN AGRICULTURAL UNIVERSITY
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  • Description
  • Claims
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Problems solved by technology

[0009] The present invention overcomes the disadvantages of single composition and single biological function of the synthetic hydrogel; overcomes the adhesion inhibition of traditional cell culture; Gel, the preparation method of the present invention is to decellularize the skin, and through matrix extraction and dialysis, to obtain an extract that is an aqueous solution at 4°C and forms a gel at body temperature

Method used

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  • Method for extracting matrix gel from cattle and donkey skin
  • Method for extracting matrix gel from cattle and donkey skin
  • Method for extracting matrix gel from cattle and donkey skin

Examples

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Embodiment Construction

[0032] 1. Gel test of the extract

[0033]Transfer the extract to a 1.5mL centrifuge tube, mark it, and store it in a refrigerator at 20°C for 48 hours. Take out the frozen extract sample and thaw it in a refrigerator at 4°C for 4h. After the sample is completely thawed, put it in a 37°C constant temperature water bath and incubate for 2-5min, and observe whether the sample to be tested forms a gel.

[0034] 2. Gradient Gel Test of Extracts

[0035] Place the skin extract on ice, observe whether there are impurities in the extract (precipitation may be drawn when the supernatant is taken after centrifugation), and perform a brief low-speed centrifugation on the extract. The extract was serially diluted with pre-cooled DMEM medium in an ice box, so that the concentration gradient of the extract was stock solution, 50%, 33.3%, 25% and DMEM medium, and placed in 37°C after pipetting evenly In the water bath, observe the time of gel formation, the state of gel formation at each...

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Abstract

The invention discloses a method for extracting matrix gel from cattle and donkey skin, and relates to the technical field of biological materials. The matrix gel is a soluble basement membrane matrix extracted from an extracellular matrix and can be used as a biological scaffold material for three-dimensional cell culture; and the matrix gel for three-dimensional cell culture can better simulate the in-vivo growth environment of cells, so that the cells can better exert the functions and characteristics of the cells. The matrix gel is characterized by being an aqueous solution at 4 DEG C and being a gel at 37 DEG C. The invention belongs to the technical field of biology; in a test, the skin of fetal cattle of 3-6 months old is taken as a research object; matrix gel is extracted from the skin; and the purpose is to establish and verify the effect of the extracted gel on the aspect of three-dimensional cell culture and screen out a more excellent biological material for three-dimensional culture of cells and organoids.

Description

technical field [0001] The invention relates to the technical field of biomaterials, in particular to a method for extracting matrix gel from cow and donkey skin. Background technique [0002] The in vitro culture of animal cells provides a clear platform for studying the physiology and pathophysiology of cells and tissues in vitro, and for cultivating replacement tissues for regenerative medicine. Traditionally, this has been accomplished by culturing single-cell populations on two-dimensional (2D) substrates, however, it has been shown that cells behave more naturally when cultured in a three-dimensional environment. Research in cell biology often relies on tissue culture, yet artificial substrates such as plastic or glass can force cells to adapt to flat and hard surfaces, distorting research results. In contrast, the true matrix of most cells in living organisms is the extracellular matrix, whose molecular composition is three-dimensional, complex, and dynamic, with var...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00A61K8/98A61Q19/00
CPCC12N5/0062A61K8/985A61Q19/00C12N2513/00C12N2533/90A61K2800/805
Inventor 张志平贺来增张君涛邓立新赵英博石亚珂王新卫董海聚
Owner HENAN AGRICULTURAL UNIVERSITY
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