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Bionic collagen membrane coated sheet and preparation method thereof

A biomimetic glue, membrane-encapsulated technology, applied in the field of cell or tissue culture materials, can solve the problems of complicated operation, low collagen coating rate, and high collagen concentration requirements, and achieve the effects of good activity, good cell affinity, and easy operation.

Pending Publication Date: 2021-09-17
SOUTH CHINA UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The National Institute of Standards and Technology prepared collagen fiber coatings on the surface of non-TC-treated cell culture well plates (polystyrene), but most of the collagen was washed away, so the concentration of collagen used when coating the well plates required high , the coating rate of collagen is low
[0006] Earlier CN103060192B reported a culture vessel coated with bionic collagen film and its preparation method, but this method of coating the surface of the vessel requires multiple washing processes, the operation is relatively cumbersome, and the coating form is limited to the culture vessel, which has certain limitations. limitation

Method used

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  • Bionic collagen membrane coated sheet and preparation method thereof
  • Bionic collagen membrane coated sheet and preparation method thereof
  • Bionic collagen membrane coated sheet and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] A method for bionic collagen film coating sheet, the steps are as follows:

[0047] (1) Type I collagen was added into 0.02M acetic acid solution to prepare a collagen acetic acid solution with a type I collagen concentration of 0.2 mg / ml.

[0048] (2) preparation of collagen reconstitution buffer; this buffer contains 270mM NaCl, 60mM HEPES, 60mM Na 2 HPO 4 .

[0049](3) Mix the collagen acetic acid solution described in step (1) and the collagen recombination buffer solution described in step (2) in a ratio of 1:1 to obtain a mixed solution, and use 1M NaOH to adjust the pH of the mixed solution to be neutral, Take 1ml of the pH-adjusted mixed solution and add it to a 12-well plate, vertically erect a polystyrene sheet in the well plate, cover the lid, and keep warm in an oven at 30 degrees Celsius for 1 hour.

[0050] (4) Open the lid of the 12-well plate containing recombinant collagen obtained in step (3), and bake in an oven at 30 degrees Celsius for 2 hours to...

Embodiment 2

[0055] A method for bionic collagen film coating sheet, the steps are as follows:

[0056] (1) Type I collagen was added into 0.005M acetic acid solution to prepare a collagen acetic acid solution with a type I collagen concentration of 0.05 mg / ml.

[0057] (2) preparation of collagen reconstitution buffer; this buffer contains 270mM NaCl, 60mM HEPES, 60mM Na 2 HPO 4 .

[0058] (3) Mix the collagen acetic acid solution described in step (1) and the buffer solution described in step (2) in a ratio of 1:1 to obtain a mixed solution, use 1M NaOH to adjust the pH of the mixed solution to be neutral, take 1ml The pH-adjusted mixed solution was added to a 12-well plate, and a polyethylene sheet was vertically erected in the well plate, covered with a lid, and kept in an oven at 30 degrees Celsius for 2 hours.

[0059] (4) Open the lid of the 12-well plate containing recombinant collagen obtained in step (3), and bake in an oven at 30 degrees Celsius for 12 hours to obtain a biomi...

Embodiment 3

[0063] A method for bionic collagen film coating sheet, the steps are as follows:

[0064] (1) Type I collagen was added into 0.05M acetic acid solution to prepare a collagen acetic acid solution with a type I collagen concentration of 0.5 mg / ml.

[0065] (2) preparation of collagen recombination buffer; this buffer contains 540mM NaCl, 120mM HEPES, 120mM Na 2 HPO 4 .

[0066] (3) Mix the collagen acetic acid solution described in step (1) and the buffer solution described in step (2) uniformly in a ratio of 3:1 to obtain a mixed solution, use 1MNaOH to adjust the pH of the mixed solution to be neutral, take Add 1ml of the pH-adjusted mixed solution into a 12-well plate, place the silicon wafer vertically in the well plate, cover the lid, and keep the temperature in an oven at 30 degrees Celsius for 5 hours.

[0067] (4) Open the lid of the 12-well plate containing recombinant collagen obtained in step (3), and dry it in an oven at 35 degrees Celsius for 48 hours until the ...

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Abstract

The invention discloses a bionic collagen membrane coated sheet and a preparation method thereof. The method comprises the following steps of preparing a collagen recombination buffer solution, uniformly mixing an acetic acid solution of collagen with the collagen recombination buffer solution to obtain a mixed solution, and adjusting the pH value of the mixed solution to be neutral; and vertically soaking a carrier in the mixed solution, carrying out heat preservation treatment, and then drying to obtain the bionic collagen membrane coated sheet. The bionic collagen membrane coated sheet obtained by a simple collagen self-assembly preparation method has the following advantages that the collagen and a carrier sheet are tightly attached, the flatness is good, the operation is simple and convenient, and the bionic collagen membrane coated sheet has a nanofiber structure and a unique 67 nm periodic thread structure in the collagen body.

Description

technical field [0001] The invention belongs to the field of cell or tissue culture materials, and in particular relates to a bionic collagen film-coated sheet and a preparation method thereof. Background technique [0002] Culturing cells on extracellular matrices helps maintain the differentiated phenotype of cells in vitro and allows the use of serum-free media for culturing cells. Currently, extracellular matrix materials commonly used in cell culture include various types of collagen, fibronectin, and laminin. As the most abundant extracellular matrix component in humans and animals, collagen has long been known for its excellent biocompatibility and biological activity. [0003] In vivo, after procollagen molecules are synthesized, the telopeptides at both ends are cut off by enzymes to form collagen molecules with a length of about 300nm and a diameter of 1.5nm, and then multiple collagen molecules self-assemble to form nano-collagen fibers with a typical D-band stru...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/00C12N2533/54
Inventor 翟志臣郝丽静石志锋卞钲淇薛永业
Owner SOUTH CHINA UNIV OF TECH
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