Preparation method of leucoderma melanocyte transplanting coating carrier system

A melanocyte and carrier system technology is applied in the field of preparation of vitiligo melanocyte transplantation coating carrier system to achieve the effects of solving the easy loss of cell suspension, reducing the loss and improving the implantation rate

Inactive Publication Date: 2013-12-04
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 of leucoderma melanocyte transplanting coating carrier system
  • Preparation method of leucoderma melanocyte transplanting coating carrier system
  • Preparation method of leucoderma melanocyte transplanting coating carrier system

Examples

Experimental program
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Effect test

Embodiment 1

[0025] (1) Preparation of melanocyte culture medium:

[0026] 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.

[0027] (2) In vitro culture of melanocytes:

[0028] 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.

[0029] Stop digestion with fetal bovine serum (1:1 volume ratio) or medium containing serum, pipette to...

Embodiment 2

[0035] (1) Preparation of melanocyte culture medium:

[0036] 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.

[0037] (2) In vitro culture of melanocytes:

[0038] 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.

[0039]Use fetal bovine serum (1:1 volume ratio) or serum-containing medium to stop digestion, pipette to obtain cel...

Embodiment 3

[0045] (1) Preparation of melanocyte culture medium:

[0046] 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.

[0047] (2) In vitro culture of melanocytes:

[0048] 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.

[0049] 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 method for preparing a coating carrier system for vitiligo melanocyte transplantation. , epinephrine and vitamin C mixed solution, pentamycin to configure melanocyte culture medium for the in vitro culture of autologous melanocytes, and use Carbomer gel and the above melanocyte culture medium to form a coating Cloth carrier system. The present invention uses the melanocyte culture medium containing human granulocyte colony-stimulating factor G-CSF to make the melanocyte proliferate in large quantities in vitro; The problem of easy loss of cell suspension during cell transplantation can reduce the loss of melanocytes, increase the implantation rate without causing damage to melanocytes, and reduce the cost of surgery. Treatment offers new avenues.

Description

Technical field [0001] The invention is a biomedical field, especially the preparation methods and applications of a vitiligo melanocyte transplantation carrier system. Background technique [0002] Vitiligo is a common dislocation of the cure skin pigmentation, and tissue pathological is mainly manifested by the lack of melanocytes.Although vitiligo has no serious impact on health, patients often have a heavy psychological burden on the appearance image, affecting normal life such as work and learning, and easily developing into psychological diseases.Traditional treatment methods include drug treatment, physical therapy and surgery. Due to the efficiency of drugs and physical therapy, surgery often becomes the final choice for patients with vitiligo.Common methods include: autologous epidermal transplantation, melanocyte suspension transplantation, etc. Although these therapies have certain effects, autologous epidermal transplantation is only applicable to the treatment of sm...

Claims

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Application Information

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Patent Type & Authority Patents(China)
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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