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Heterologous or heterogenic decelled epidermis substitute used for human fibroblast-like cell modification

A human fibroblast, acellular dermis technology, applied in prosthesis, medical science and other directions, can solve the problems of unfavorable epidermal cell adhesion and proliferation, slow vascularization, low affinity, etc. Transplantation, vascularization promotion, slow proliferation effect

Inactive Publication Date: 2006-11-29
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Fibroblasts secrete a variety of cytokines, growth factors and extracellular matrix components during culture, such as interleukin-6, interleukin-8, transforming growth factor-β, keratinocyte growth factor, acidic fibroblast growth Factors, basic fibroblast growth factor, laminin, hyaluronic acid, collagen type IV, etc., make the acellular dermal substitute have higher biological activity, improve affinity, and be suitable for epidermal cell adhesion and growth and proliferation, thereby overcoming the defects of commonly used acellular dermal substitutes such as low affinity, slow vascularization, and unfavorable adhesion and proliferation of epidermal cells

Method used

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Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0010] The preparation of dermis substitute of the present invention, if use heterogeneous decellularized dermis substitute, then can press 3-4 * 10 on its surface 5 piece / cm 2 Inoculate human fibroblasts pretreated with mitomycin C at a cell density, and culture them in vitro for 1-2 weeks. The heterogeneous acellular dermal substitutes containing monolayer or stratified fibroblasts can be placed in cell culture Reserve in liquid.

[0011] If allogeneic acellular dermal substitute is used, press 2-3×10 5 piece / cm 2 Cell density inoculation of human fibroblasts pretreated with mitomycin C, after 1-2 weeks of in vitro culture, the formed allogeneic acellular dermal substitute containing monolayer or stratified fibroblasts was placed in cell culture medium spare.

Embodiment 1

[0012] Example 1. Preparation of allogeneic acellular dermal substitute modified by human fibroblasts

[0013] 1. Preparation of allogeneic acellular dermal substitute:

[0014] Prepare the allogeneic acellular dermal substitute with a thickness of 0.4mm by conventional enzyme digestion and detergent soaking method;

[0015] 2. Inoculation of human fibroblasts:

[0016] The human fibroblasts cultured to the third generation were routinely added to the cell culture medium, and the cell concentration was 2×10 6 cells / ml, then add mitomycin C to make the final concentration 10 μg / ml, incubate at 37°C for about 3 hours, and then add 2×10 5 piece / cm 2 The cell density is seeded on the surface of the allogeneic decellularized dermal substitute, placed in DMEM culture medium and cultured for 10 days to form the allogeneic decellularized dermal substitute modified by stratified human fibroblasts.

Embodiment 2

[0017] Example 2. Preparation of human fibroblast-modified xenogeneic acellular dermal substitute

[0018] 1. Preparation of xenogeneic acellular dermal substitute:

[0019] The xenogeneic acellular dermis substitute was prepared by conventional enzyme digestion and detergent soaking method with the split pigskin, with a thickness of 0.3 mm;

[0020] 2. Inoculation of human fibroblasts:

[0021] Human fibroblasts cultured to the fifth generation were added to the DMEM medium, and the cell concentration was 2×10 6 cells / ml, then add mitomycin C to make the final concentration 10 μg / ml, incubate at 37°C for about 3 hours, and then add 4×10 5 piece / cm 2 The cell density was seeded on the surface of the heterogeneous decellularized dermal substitute, placed in DMEM medium and cultured for 7 days to form a heterogeneous decellularized dermal substitute modified with a single layer of human fibroblasts.

[0022] The method for preparing composite skin with the decellularized der...

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Abstract

A human fibroblast modified foreign cell-removed epidermis substitute with high bioactivity and affinity is prepared through treating the human fibroblasts by mitomycin C, inoculating them on the surface of foreign cell-removed epidermis substitute, and external culture to become the epidermis substitute containing single or double layers of human fibroblasts.

Description

technical field [0001] The invention relates to the technical field of medical wound repair, in particular to a human fibroblast-modified heterogeneous or heterogeneous decellularized dermis substitute used as a dermal scaffold or an epidermal cell culture carrier. Background technique [0002] Recently, allogeneic or heterogeneous acellular dermal substitutes, as a dermal scaffold, have been initially applied in the repair of deep skin defect wounds and scar plastic surgery. On the other hand, the acellular dermal substitute can also be used as a carrier for epidermal cell culture, and after constructing a composite skin in vitro, it can repair full-thickness skin defect wounds at one time. Allogeneic or heterogeneous acellular dermal substitutes need to be treated with enzyme digestion and detergents during the preparation process, because while removing antigenic cellular components, some extracellular matrix such as laminin, hyaluronic acid, type IV...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/38A61L27/60
Inventor 肖仕初夏照帆朱世辉王广庆田建广韩姝
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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