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Preparation method of fish scale tissue scaffold for cell culture

A fish scale tissue and cell culture technology, applied in general culture methods, cell culture supports/coatings, biochemical equipment and methods, etc., can solve the problems of inability to guide cell growth and low cell affinity.

Pending Publication Date: 2021-12-28
OCEAN UNIV OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] It has been reported in the literature that some artificial cell scaffold materials have low affinity to cells and cannot guide cell growth

Method used

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  • Preparation method of fish scale tissue scaffold for cell culture
  • Preparation method of fish scale tissue scaffold for cell culture
  • Preparation method of fish scale tissue scaffold for cell culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Take 3 fish scales, add 1.5 times volume containing benzylsulfonyl fluoride (0.35 mL / L) Tris buffer (10 mm, pH = 7.4) to stir 42 hours to sample 1; 0.5% Triton X-100 solution Stirring 42 hours with benzylsulfonyl flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu. 2; Sample 3 was obtained; for 36 hours with 0.5% Triton X-100 Tris buffer (0.05 m), it was soaked in the PBS solution for 48 hours, and sample 4 was obtained, so soaking with acetic acid (...

Embodiment 2

[0029] Three fish scales were taken, plus 1x volume containing benzylsulfonyl flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu. Tris buffer (0.05 ml / L) containing benzylsulfonyl flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu flu ...

Embodiment 3

[0031] Take 3 fish scales, plus 2 times volume containing benzylsulfonyl fluoride (0.35 mL / L) Tris buffer (10 mm, pH = 7.4) to stir 40 hours to sample 1; 1% Triton X-100 Solution with Tris buffer containing benzylsulfonyl fluoride (0.35 ml / L) was stirred for 40 hours of sample 2; DNase (5 μg / mL) and RNase (5 μg / ml) at 37 ° C Digestion 2 hours sample 3; after 30 hours with 1% Triton X-100 Tris buffer (0.05 m), soak for 64 hours in PBS solution, sample 4; soaking with acetic acid (0.2 m) for 1 hour Sample HCS ;; Stirring of 15% by mass concentration of citric acid 7 hours to decide samples CS1; stirring at a mass concentration of 45% citric acid 2.5 hours quadly decoction sample CS2; for HCS, CS1, CS2 75% alcohol and Ultraviolet irradiation and BSA solution (200 μg / ml, pH = 7.4) were mixed according to a mass volume ratio of 1: 6, and 37 ° C were incubated for 12 hours and soaked in 0.3 mm of Jingni PBS buffer (pH = 7.4) In the treatment of 40 minutes, it is neutralized wi...

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Abstract

The invention discloses a preparation method of a fish scale tissue scaffold for cell culture. The preparation method comprises the following steps: taking fish scales as raw materials, respectively carrying out decellularization, primary decalcification, secondary decalcification treatment, 75% alcohol soaking and ultraviolet radiation sterilization on the fish scales, adsorbing bovine serum albumin, and then carrying out genipin curing treatment to obtain three fish scale scaffold materials used for adherent culture of cells. The fish scale cell scaffold materials disclosed by the invention have good cell affinity and capability of guiding cells to migrate and grow directionally, and are beneficial to quickly repairing cell defects, so that the fish scale scaffold disclosed by the invention has a good application prospect in the fields of cell culture and tissue engineering.

Description

Technical field [0001] The present invention belongs to a method for preparing a fish scale tissue bracket that can be used in cell culture in the field of tissue engineering. Background technique [0002] As a large country of freshwater fish, China has a large amount of fish scales, which is a low price and easy to obtain biological resources. Document analysis showed that there were two different layers of fish scales: the outer bone layer consists of hydroxyapatite crystals, the inner or bottom layer consists of a ply-like-forming plate structure containing collagen fibers, and thereof is similar to the osteocellular matrix. In addition, the main components of the fish scales, hydroxyapatite and type I collagen fibers become research hotspots in the field of biological resources, both of which exhibit good biological characteristics. Type I collagen is the main component of extracellular matrix, and has functions such as mechanical protection or physiological adjustment of th...

Claims

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Application Information

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IPC IPC(8): C12N5/00C12N5/0775
CPCC12N5/0062C12N5/0663C12N5/0068C12N2513/00C12N2537/10C12N2533/50C12N2533/90
Inventor 不公告发明人
Owner OCEAN UNIV OF CHINA