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A method for culturing human melanocytes based on an asymmetric membrane

A technology for cell culture and melanocytes, which is applied in the field of biomedicine, can solve problems that have not been involved in the application of melanocyte culture, melanocyte culture, etc., to promote growth and maintain activity, maintain cell activity, and be easy to operate Effect

Active Publication Date: 2017-03-08
HANGZHOU THIRD HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN102648987A discloses a sodium hyaluronate-chitosan composite material prepared as an asymmetric surface material with a smooth surface and a spongy surface to promote the growth of epithelial cells and endothelial cells, but the material is not used for the cultivation of melanocytes
The relevant published patents of existing asymmetric membranes are mainly used for wound dressings, and do not involve the application in melanocyte culture

Method used

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  • A method for culturing human melanocytes based on an asymmetric membrane
  • A method for culturing human melanocytes based on an asymmetric membrane
  • A method for culturing human melanocytes based on an asymmetric membrane

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1. Preparation of chitosan asymmetric membrane material

[0036] A, chitosan is dissolved with the acetic acid solution of 1M, is made into the chitosan solution that concentration is 1.5%;

[0037] b. Inject 2.5ml of chitosan solution into the cell culture plate, place the cell culture plate in a blast oven, and dry it at 65°C for 0.5 hours;

[0038] C, add sodium hydroxide, sodium carbonate and water and be the sodium hydroxide-sodium carbonate aqueous solution of 2:1:47 with weight ratio, soak 24 hours, make dense membrane lower floor solution phase-separate, form porous structure;

[0039] d. Wash with deionized water until the pH of the solution is neutral;

[0040] e. Freeze-dry at -55ºC for 12 hours to obtain an asymmetric membrane;

[0041] f. Place the dried chitosan asymmetric membrane in 0.5M Na 2 SO 4 Soak in the solution for 30 minutes, then rinse with plenty of pure water;

[0042] g. Place the cell culture plate covered with the chitosan a...

Embodiment 2

[0044] Embodiment 2. Preparation of chitosan-gelatin-based composite asymmetric membrane material

[0045] A, chitosan is dissolved with the acetic acid solution of 1M, is made into the chitosan solution that concentration is 1.5%;

[0046] b. Dissolve the gelatin with purified water to make a 1% gelatin solution;

[0047] C, chitosan and gelatin are mixed with the weight ratio of 20:1, stir, obtain chitosan-gelatin mixed solution;

[0048] d. Inject 2.5 ml of the mixed solution into the cell culture plate, place the cell culture plate in a blast oven, and dry it at 35ºC for 4 hours;

[0049] e, add sodium hydroxide, sodium carbonate and water and be the sodium hydroxide-sodium carbonate aqueous solution of 2:1:47 with weight ratio, soak for 24 hours, make dense membrane lower layer solution phase-separate, form porous structure;

[0050] f, washing with deionized water until the pH of the solution is neutral;

[0051] g. Freeze-dry at -45ºC for 24 hours to obtain an asymme...

Embodiment 3

[0055] Embodiment 3. Preparation of chitosan-polyethylene glycol based composite asymmetric membrane material

[0056] A, chitosan is dissolved with the acetic acid solution of 1M, is made into the chitosan solution that concentration is 1.5%;

[0057] b. Dissolve polyethylene glycol with purified water to make a 1% polyethylene glycol solution;

[0058] C, chitosan and polyethylene glycol are mixed with the weight ratio of 20:1, stir, obtain chitosan-polyethylene glycol mixed solution;

[0059] d. Inject 2.5 ml of the mixed solution into the cell culture plate, place the cell culture plate in a blast oven, and dry it at 35ºC for 4 hours;

[0060] e, add sodium hydroxide, sodium carbonate and water and be the sodium hydroxide-sodium carbonate aqueous solution of 2:1:47 with weight ratio, soak for 24 hours, make dense membrane lower layer solution phase-separate, form porous structure;

[0061] f, washing with deionized water until the pH of the solution is neutral;

[0062]...

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Abstract

The invention discloses a method for culturing human melanocyte based on an asymmetric membrane. According to the method, an asymmetric membrane with excellent biocompatibility is constructed by utilizing the tissue engineering technology, and the melanocyte or melanocyte, fibroblast and keratinocyte are cultured and transferred or co-cultured and co-transferred. The method is suitable for adjusting depigmentation, such as leukoderma, and skin color adjustment. The invention relates to the preparation of the asymmetric membrane and construction of a cell-asymmetric membrane composite material. The asymmetric structure is introduced into the membrane material, so that cell culture and activity maintenance can be promoted.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a method for culturing melanocytes based on asymmetric biofilm materials. [0002] Background of the invention [0003] Vitiligo is a common skin depigmentation skin disease with an incidence rate of 0.5-2% in the population. The cause is the loss of function of melanocytes in human skin. Vitiligo is manifested as partial or generalized leukoplakia. Due to the lack of melanin in the leukoplakia, it is easy to be burned by the sun and even cause cancer. At the same time, it can also bring social anxiety to patients and cause mental illness. [0004] There are currently three main types of therapy for the treatment of vitiligo: drug therapy, photochemotherapy, and surgical therapy. Drug therapy and photochemotherapy have general curative effects and severe side effects, so people pay more attention to surgical therapy. Surgical treatment uses tissue engineering technology to...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/071A61L27/38A61L27/24A61L27/22A61L27/20A61L27/18A61L27/60
Inventor 许爱娥王文俊邹洁辉蒋森阳尉晓冬
Owner HANGZHOU THIRD HOSPITAL
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