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Preparation method and application of coating carrier system for vitiligo melanocyte transplantation

A melanocyte and carrier system technology is applied in the field of preparation of vitiligo melanocyte transplantation coating carrier system, which can solve the problems of easy loss of cell suspension, reduce the operation cost, solve the problem of easy loss of cell suspension, and reduce the loss. amount of effect

Inactive Publication Date: 2011-12-07
JIANGSU PROVINCE HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The applicant has carried out research and improvement on the above-mentioned problems, and provides a preparation method and application of a coating carrier system for vitiligo melanocyte transplantation, and configures a Excellent melanocyte culture medium, so that melanocytes can proliferate in large quantities in vitro; and solve the problem of easy loss of cell suspension during the existing cell transplantation process, improve the implantation rate and reduce the operation cost

Method used

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  • Preparation method and application of coating carrier system for vitiligo melanocyte transplantation
  • Preparation method and application of coating carrier system for vitiligo melanocyte transplantation
  • Preparation method and application of coating carrier system for vitiligo melanocyte transplantation

Examples

Experimental program
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Effect test

Embodiment 1

[0023] (1) Preparation of melanocyte culture medium:

[0024] Take 100 milliliters (mL) of melanocyte basal culture medium (Hams F12), add 10 mL of fetal bovine serum (FBS), 200 microliters (μL) of human granulocyte colony-stimulating factor G-CSF (trade name: Ruibai), the concentration 100 μL of melanopoietin α-MSH at 100 nanograms / milliliter (ng / mL), 100 μL of isomethylxanthine IBMX, 1.0 mL of a mixture of adrenaline and vitamin C at a concentration of 1.5 μg / mL (μg / mL) , 1.0 mL of penicillin-streptomycin solution, stored in a refrigerator at 4°C.

[0025] (2) In vitro culture of melanocytes:

[0026] The skin donor area was routinely disinfected, and autologous skin samples were obtained under sterile conditions; after washing with phosphate buffered saline (PBS), 9 mL of 0.25% trypsin-EDTA digestion solution was added, and digested in a 37°C incubator for 14 minutes.

[0027] Stop digestion with fetal bovine serum (1:1 volume ratio) or medium containing serum, pipette to...

Embodiment 2

[0033] (1) Preparation of melanocyte culture medium:

[0034] Take 120 milliliters (mL) of melanocyte basal culture medium (Hams F12), add 12 mL of fetal bovine serum (FBS), and 170 microliters (μL) of human granulocyte colony-stimulating factor G-CSF (trade name: Ruibai), the concentration Melanopoietin α-MSH 120 μL at 100 nanograms / ml (ng / mL), isomethylxanthine IBMX 110 μL, a mixture of epinephrine and vitamin C at a concentration of 1.5 μg / ml (μg / mL) 0.8 mL , 1.2 mL of penicillin-streptomycin solution, stored in a refrigerator at 4°C.

[0035] (2) In vitro culture of melanocytes:

[0036] The skin donor area was routinely disinfected, and autologous skin samples were obtained under sterile conditions; after washing with phosphate buffered saline (PBS), 8 mL of 0.25% trypsin-EDTA digestion solution was added, and digested in a 37°C incubator for 10 minutes.

[0037] Use fetal bovine serum (1:1 volume ratio) or serum-containing medium to stop digestion, pipette to obtain ce...

Embodiment 3

[0043] (1) Preparation of melanocyte culture medium:

[0044] Take 80 milliliters (mL) of melanocyte basal culture medium (Hams F12), add 8 mL of fetal bovine serum (FBS), 230 microliters (μL) of human granulocyte colony-stimulating factor G-CSF (trade name: Ruibai), the concentration Melanopoietin α-MSH 80 μL at 100 nanograms / milliliter (ng / mL), isomethylxanthine IBMX 90 μL, a mixture of epinephrine and vitamin C at a concentration of 1.5 μg / mL (μg / mL) 1.2 mL , 0.8 mL of penicillin-streptomycin solution, stored in a refrigerator at 4°C.

[0045] (2) In vitro culture of melanocytes:

[0046] The skin donor area was routinely disinfected, and autologous skin samples were obtained under sterile conditions; after washing with phosphate buffered saline (PBS), 10 mL of 0.25% trypsin-EDTA digestion solution was added, and digested in a 37°C incubator for 15 minutes.

[0047] Use fetal bovine serum (1:1 volume ratio) or serum-containing medium to stop digestion, pipette to obtain c...

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Abstract

The invention relates to a preparation method of a leucoderma melanocyte transplanting coating carrier system. The method comprises the following steps of: preparing a melanocyte culture medium from a melanocyte basic culture solution Hams F12, fetal calf serum, a human granulocyte colony stimulating factor (G-CSF), a melanogenesis promoter, iso-methyl xanthine, adrenalin, a vitamin C mixed liquor and penicillin streptomycin for in-vitro culturing of autologous melanocytes; and constructing a coating carrier system by using Carbomer gel and the melanocyte culture medium. In the invention, the melanocyte culture medium containing the human G-CSF is used, so that massive in-vitro proliferation of melanocytes is realized; and the Carbomer gel plays a good role in fixing, so that the problem of easy loss of a cell suspension in the conventional melanocyte transplanting process is solved, the loss amount of melanocytes is reduced, the implantation rate is increased, damage to the melanocytes is avoided, the surgery cost is reduced, the preparation method is easy and safe for operating during clinical application, and a new way is provided for the treatment of leucoderma.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a preparation method and application of a coating carrier system for transplantation of vitiligo melanocytes. Background technique [0002] Vitiligo is a common refractory skin depigmentation disease, histopathologically mainly manifested as the loss of melanocytes. Although vitiligo has no serious impact on health, patients often have a heavy psychological burden for appearance and image, which affects normal life such as work and study, and easily develops into mental illness. Traditional treatment methods include drug therapy, physical therapy, and surgery. Due to the limitation of the effectiveness of drugs and physical therapy, surgery is often the final choice for patients with vitiligo. Commonly used methods include: autologous epidermal transplantation, melanocyte suspension transplantation, etc. Although these treatments have certain curative effects, autologous e...

Claims

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Application Information

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IPC IPC(8): A61L27/38C12N5/071
Inventor 鲁严吴迪朱文元李雪周梅华
Owner JIANGSU PROVINCE HOSPITAL
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