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Artificial skin and preparation method thereof

A technology for artificial skin and dermis, applied in the field of biomedicine, can solve the problems of difficult to form a smooth surface, poor mechanical properties, uneven application, etc., and achieve the effects of fast skin healing, good mechanical properties, and smooth wounds

Active Publication Date: 2015-04-29
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the first problem facing the use of ADSCs in the preparation of wound repair materials is differentiation.
In the prior art, there are few studies on the epidermal cell differentiation of ADSCs and the fibroblast differentiation of ADSCs. The only research results show that the time required for ADSCs to differentiate into epidermal cells is about 20 days, which is relatively long.
[0005] In addition, in the prior art, most of the wound repair materials that use stem cells as raw materials are directly applied to the wound surface. It is difficult to form a smooth surface during the healing process, and the problem of uneven application is inevitable during the application process.
Mixing cells and scaffold materials to make artificial skin is an effective means to solve the problems in the application process. However, most of the existing artificial skins are fragile, have poor mechanical properties, and are prone to breakage during use.

Method used

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  • Artificial skin and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0106] Example 1 Isolation and cultivation of human adipose stem cells (ADSCs)

[0107] The human abdominal subcutaneous adipose tissue extract obtained through liposuction (endocrine diseases and infectious diseases have been excluded before the operation) was centrifuged at 800g, rinsed with PBS buffer, soaked, and washed 3 times at 800gx4min.

[0108] Remove visible blood vessels and hoof tissue, use ophthalmic scissors to fully cut them into 50ml centrifuge tubes, add 2 times the volume of 0.5% collagenase, 1 times the volume of 1% BSA and double antibodies, mix well, seal, and put Digest in a shaker at 37°C for 50 minutes until it becomes a paste.

[0109] Add an equal volume of low-sugar DMEM containing 10% fetal bovine serum to stop digestion, centrifuge at 1800r / min for 10min, and divide into 3 layers after centrifugation, the upper layer is oil and undigested adipose tissue, the middle layer is supernatant, and the lower layer is It is a pellet of mixed cells such as...

Embodiment 2

[0115] Example 2 Identification of surface markers of human adipose stem cells (ADSCs)

[0116] Take the cells of the third generation in the logarithmic growth phase, suck out the medium, add 0.25% trypsin + 0.02% EDTA digestion solution for digestion, then stop the digestion with an appropriate amount of serum, and gently pipette into a single cell suspension.

[0117] Centrifuge at 1000rpm for 10min, discard the supernatant, wash the cells 3 times with cold PBS at 4°C and resuspend evenly, adjust the cell concentration to about 10 5 -10 6 pieces / ml.

[0118] Take 2 loading tubes and add 500ul single cell suspension to each tube. After centrifugation, tube 1 is marked as the standard control, and tube 2 is added with 2μl FITC or PE-labeled mouse anti-human cell surface molecules CD59, CD45, CD34, CD105 Antibody working solution.

[0119] At room temperature, protected from light, incubate for 20 minutes.

[0120] Wash twice with PBS to remove unbound antibodies, resuspen...

Embodiment 3

[0122] Example 3 Induced Differentiation of Human Adipose Stem Cells to Epidermal Cells

[0123] The third to fifth generation ADSCs prepared in Example 1 were taken, the culture supernatant was sucked off, and then digested by adding 0.25% trypsin + 0.02% EDTA digestion solution, and then an appropriate amount of serum was used to terminate the digestion. Centrifuge at 1200r / min for 5min, resuspend the adipose stem cells in complete medium, adjust the density to 1x104 / ml, and inoculate in 6-well plates, 2ml per well. When the cells reach 40%-50% confluence, use the medium shown in Table 1, induce culture in the same environment, replace the induction medium every 2 days, and after 7 days of culture, take some cells to detect related indicators, and use epidermal cells for the rest of the cells. The serum medium SFM continued to be cultured and passaged, and the complete medium was replaced every 2 days.

[0124] Table 1 Experimental design of induction and differentiation of...

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Abstract

The invention relates to the technical field of biomedicines, and particularly relates to an artificial skin and a preparation method thereof. The artificial skin comprises a dermal layer and an epidermal layer, wherein the dermal layer and the epidermal layer are fitted; the dermal layer comprises a timbering material and fibroblast; the epidermal layer consists of a timbering material and epidermal cells; the timbering material consists of DMEM / F12 culture media, I type mouse collagen, chitin, glycosaminoglycan and glutaraldehyde; the epidermal cells and fibroblast are produced by fat stem cell differentiation. No smearing is needed in use, the structure of the artificial skin is more suitable for the self-situations of patients, and the artificial skin can achieves a better therapeutic effect on the wound restoration, the heeling speed of the skin is faster, and the wound is more smooth after being healed. Furthermore, the artificial skin has small possibility of breaking in use due to good mechanical performance.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to an artificial skin and a preparation method thereof. Background technique [0002] As the largest organ of the body, the skin functions to prevent bacterial erosion and infection, protect the body, regulate body temperature, and excrete body fluids. The skin can be divided into three layers, namely the epidermis, dermis and subcutaneous tissue. The skin has a strong ability to regenerate. However, the repair and regeneration of large-area skin damage caused by chronic ulcers, severe burns, and severe trauma is still a technical problem in the medical field. In addition to autologous skin grafting, people have used allogeneic skin grafting and other related repair materials to cover wounds, but both autologous and allogeneic skin grafts are limited by donor sources and immune rejection. [0003] Therefore, it is urgent to find an ideal material without rejection, promoting s...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/38A61L27/24A61L27/20A61L27/00
Inventor 陈海佳王一飞葛啸虎马岩岩王小燕
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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