Method for culturing human melanocyte based on nano-fiber scaffold

A technology of nanofibers and melanocytes, applied in the field of biomedicine, to achieve the effect of promoting proliferation, good mechanical properties and maintaining activity

Active Publication Date: 2014-06-18
HANGZHOU THIRD HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The existing published nanofiber scaffold patents are mainly used for repair of tissue engineering, and there is no public report on the cultivation of human melanocytes

Method used

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  • Method for culturing human melanocyte based on nano-fiber scaffold
  • Method for culturing human melanocyte based on nano-fiber scaffold
  • Method for culturing human melanocyte based on nano-fiber scaffold

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Embodiment 1. Preparation of chitosan nanofiber scaffold material

[0026] A, chitosan is dissolved with trifluoroacetic acid / dichloromethane weight ratio 3:1 mixed solvent, is made into the chitosan solution that concentration is 5%;

[0027] b. Electrospinning is carried out at a voltage of 15KV, a propulsion rate of 1ml / h, a receiving distance of 15cm, and a temperature of 20oC;

[0028] C, the chitosan nanofiber support is baked under vacuum, 50oC for 12 hours;

[0029] d. Under 20oC, the chitosan nanofiber support was cross-linked with sodium sulfate for 2 hours, and after the cross-linking treatment, the residual cross-linking agent was rinsed with pure water;

[0030] e, soak the chitosan nanofiber support with a mass fraction of 4% NaOH solution for 24 hours, then wash it to PH=7.2-7.4 with a large amount of pure water;

[0031] f, the chitosan nanofiber support is placed in a blast oven, and dried for 24 hours at 50oC;

[0032] g, obtaining the chitosan nano...

Embodiment 2

[0033] Embodiment 2. Preparation of chitosan-gelatin nanofiber scaffold material

[0034] a. Dissolve chitosan-gelatin (weight ratio 10:1) with trifluoroethanol / dichloromethane (weight ratio 2:1) mixed solvent to form a chitosan-gelatin solution with a concentration of 4%;

[0035] b. Electrospinning is carried out at a voltage of 20KV, a propulsion rate of 3ml / h, a receiving distance of 18cm, and a temperature of 30oC;

[0036] C, the chitosan-gelatin nanofiber support is baked under vacuum, 25oC for 24 hours;

[0037] d. Under 20°C, the chitosan-gelatin nanofiber scaffold was cross-linked with sodium citrate for 12 hours, and after the cross-linking treatment, the residual cross-linking agent was rinsed with pure water;

[0038] e, with a mass fraction of 8% Na 2 CO 3 Soak the chitosan-gelatin nanofiber scaffold in the solution for 24 hours, and then wash it with a large amount of pure water to PH=7.2-7.4;

[0039] f, the chitosan-gelatin nanofiber support is placed in a b...

Embodiment 3

[0041] Embodiment 3. Preparation of chitosan-polyethylene glycol nanofiber scaffold material

[0042] a. Dissolve chitosan-polyethylene glycol (weight ratio 5:1) with dimethyl sulfoxide to make a chitosan-polyethylene glycol solution with a concentration of 6%;

[0043] b. Electrospinning is carried out at a voltage of 25KV, a propulsion rate of 3ml / h, a receiving distance of 12cm, and a temperature of 40oC;

[0044] C, the chitosan-polyethylene glycol nanofiber support is baked under vacuum, 25oC for 24 hours;

[0045] d. Under 50oC, the chitosan-polyethylene glycol nanofiber scaffold was cross-linked with glutaraldehyde for 12 hours, and after the cross-linking treatment, the residual cross-linking agent was rinsed with pure water;

[0046] e, with a mass fraction of 6% NaOH / Na 2 CO 3 (weight ratio 1:1) Soak the chitosan-polyethylene glycol nanofiber scaffold in the solution for 24 hours, and then wash it with a large amount of pure water to PH=7.2-7.4;

[0047] f, the c...

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Abstract

The invention discloses a method for culturing human melanocyte based on a nano-fiber scaffold. Melanocyte or melanocyte, fibroblast and keratinocyte can be (co-cultured) cultured and transferred by utilizing a nano-fiber scaffold which is constructed by utilizing a tissue engineering technology and has an excellent biocompatibility, and the method is applicable to depigmentation (such as leukoderma) and can be used for regulating skin color. The invention also relates to preparation of the nano-fiber scaffold and construction of a cell-nano-fiber scaffold composite material. Because the nano-fiber scaffold has a porous structure, 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 nanofiber scaffolds of biological materials. Background of the invention [0002] Vitiligo is an acquired skin depigmentation disease, manifested as localized or generalized depigmentation, characterized by the formation of leukoplakia, and histology and immunocytochemistry showed that the epidermal melanocytes in the lesions disappeared. Vitiligo has an incidence rate of 0.5-2% in the population. Due to the lack of melanin in the white spot, it is easy to be burned by the sun and even cause cancer. At the same time, it will also bring social anxiety to patients and cause mental illness. Traditional treatment methods include oral medicine, ultraviolet radiation and surgical treatment. But drug therapy and ultraviolet light exposure have limited cure rates. Surgical treatment methods mainly include autologous epidermal transplan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/20A61L27/44A61L27/36A61L27/54A61L27/56
Inventor 许爱娥王文俊蒋森阳尉晓冬
Owner HANGZHOU THIRD HOSPITAL
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