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Method for compounding bone marrow mesenchymal stem cells with human acellular dermis and application of prepared tissue engineering skin in repairing skin defects

A bone marrow mesenchymal, acellular dermis technology, applied in the field of tissue engineering skin and skin defect repair, can solve the problems of tight skin source, insufficient donor site, immune rejection and spread of diseases, and achieves good healing promotion and broad application prospects. Effect

Inactive Publication Date: 2022-03-01
杭州凤喆凰生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these methods have disadvantages such as insufficient donor area, tight skin source, immune rejection and disease transmission.

Method used

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  • Method for compounding bone marrow mesenchymal stem cells with human acellular dermis and application of prepared tissue engineering skin in repairing skin defects

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] A compound method of bone marrow mesenchymal stem cells and human decellularized dermis: comprising the following steps:

[0021] a. Put the decellularized dermis on a cell culture plate, add complete medium to soak for 18 hours, and incubate in a 37°C incubator for 1 hour;

[0022] b. Digest the third-generation bone marrow mesenchymal stem cells in the logarithmic growth phase with 0.25% trypsin solution, stop the digestion after 3 minutes, pipette the suspended cells, collect the suspension, centrifuge at 1200rpm / min for 8 minutes, and discard the supernatant; Resuspend the cells in complete medium, incubate at 37°C for 36 hours, centrifuge at 1200rpm / min for 8 minutes, discard the supernatant, and resuspend the cells again;

[0023] c by cell density 10 3 Inoculate the bone marrow mesenchymal stem cells obtained in step b on a cell culture plate containing decellularized dermis at 37°C, 5% CO 2 , cultured in a sterile incubator with 95% humidity, changing the med...

Embodiment 2

[0029] A compound method of bone marrow mesenchymal stem cells and human decellularized dermis: comprising the following steps:

[0030] a. Put the decellularized dermis on a cell culture plate, add complete medium to soak for 24 hours, and incubate in a 37°C incubator for 2 hours;

[0031] b. Digest the third-generation bone marrow mesenchymal stem cells in the logarithmic growth phase with 0.25% trypsin solution, stop the digestion after 5 minutes, pipette the suspended cells, collect the suspension, centrifuge at 1500rpm / min for 5 minutes, and discard the supernatant; Resuspend the cells in complete medium, incubate at 37°C for 60 hours, centrifuge at 1500 rpm / min for 5 minutes, discard the supernatant, and resuspend the cells again;

[0032] c by cell density 10 5 Inoculate the bone marrow mesenchymal stem cells obtained in step b on a cell culture plate containing decellularized dermis at 37°C, 5% CO 2 , cultured in a sterile incubator with 95% humidity, changing the m...

Embodiment 3

[0038] A compound method of bone marrow mesenchymal stem cells and human decellularized dermis: comprising the following steps:

[0039] a. Put the decellularized dermis on a cell culture plate, add complete medium to soak for 20 hours, and incubate in a 37°C incubator for 1.5 hours;

[0040] b. Digest the third-generation bone marrow mesenchymal stem cells in the logarithmic growth phase with 0.25% trypsin solution, stop the digestion after 4 minutes, pipette the suspended cells, collect the suspension, centrifuge at 1300rpm / min for 6 minutes, and discard the supernatant; Resuspend the cells in complete medium, incubate at 37°C for 48 hours, centrifuge at 1300rpm / min for 6 minutes, discard the supernatant, and resuspend the cells again;

[0041] c by cell density 10 4 Inoculate the bone marrow mesenchymal stem cells obtained in step b on a cell culture plate containing decellularized dermis at 37°C, 5% CO 2 , cultured in a sterile incubator with 95% humidity, changing the ...

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Abstract

The invention discloses a method for compounding mesenchymal stem cells with human acellular dermis and application of prepared tissue engineering skin in repairing skin defects, and belongs to the technical field of biology. The mesenchymal stem cell composite human decellularized dermis prepared by adopting the method for compounding the mesenchymal stem cells with the human decellularized dermis has a very good healing promoting effect on skin defect wounds, especially on refractory wounds such as large-area deep burns, burn residual wounds, pressure sores, sclerotin and tendon defects, and has a very good healing promoting effect on the skin defect wounds. And after tumor resection, chronic ulcer wounds and the like also have a certain effect of promoting healing, so that the preparation has a wide application prospect.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for combining bone marrow mesenchymal stem cells with human decellularized dermis and the use of the prepared tissue engineered skin in repairing skin defects. Background technique [0002] As the natural protective barrier of the human body, the skin plays an important role in maintaining the stability of the internal environment and preventing the invasion of external microorganisms. Skin defects caused by various reasons are very common in clinical practice. At present, the most commonly used treatment methods are autologous skin transplantation, allogeneic skin or xenograft skin transplantation, and composite skin transplantation. However, these methods have disadvantages such as insufficient donor site, tight skin source, immune rejection and disease transmission. Therefore, it is particularly necessary to find better skin substitutes to treat skin defects. [0003] W...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/60A61L27/38A61L27/36C12N5/0775
CPCA61L27/60A61L27/3834A61L27/362A61L27/3687C12N5/0663A61L2430/40C12N2509/00
Inventor 季子云李杰
Owner 杭州凤喆凰生物科技有限公司