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Method for constructing tissue engineering double-layered skin and the application thereof

A tissue engineering and construction method technology, applied in medical science, prosthesis, etc., can solve the problems of no self-proliferation and differentiation, pathogenic microorganism infection, etc., and achieve the effect of huge market value potential, accelerated healing, and broad application prospects

Inactive Publication Date: 2007-10-31
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although Apligraft TM The purpose of reconstructing the dermis and epidermis at the same time was achieved in one surgical operation, but due to the use of allogeneic cells as the source of seed cells, there is an inevitable immune rejection and the risk of potential pathogenic microorganism infection; and the seed cells used are all Terminally differentiated mature cells without the ability to self-proliferate and differentiate

Method used

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  • Method for constructing tissue engineering double-layered skin and the application thereof
  • Method for constructing tissue engineering double-layered skin and the application thereof
  • Method for constructing tissue engineering double-layered skin and the application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1. Taking bone marrow MSC as an example, stem cells from different tissue sources were isolated in vitro

[0034] Extract the bone marrow directly from the patient under aseptic conditions, add an equal amount of normal saline to dilute, filter through a 200-mesh sieve, add 6% HES (B. Cell suspension was separated (1000g × 20min) with Percoll separation medium (Amersham Biosciences, product number 17-0891-01) with a specific gravity of 1.073g / ml, and the middle layer cells were carefully sucked out with 10% fetal bovine serum (Biochrom, Inc., Product No. S0115) in low-sugar DMEM medium (Sigma Company, product No. D-5523) was resuspended, and inoculated in T-25 culture flasks. Set CO 2 Incubator (temperature set at 37°C, CO 2 The concentration is 5%); culture in 72h; change the medium after culturing for 72 hours, and then change the medium for the culture system according to the cell growth rate. When the cells in the culture bottle grow to more than 80% confl...

Embodiment 2

[0035] Embodiment 2, in vitro induction of MSC differentiation to epidermal cells

[0036] The bone marrow-derived MSCs were planted on a well plate pre-coated with Matrigel gel (BD company, product number 354240), and the low-sugar DMEM medium (Sigma company, product number D-5523) containing (Biochrom company, product number S0115) was added. °C, 5% CO 2 Cultivate in the incubator until the cells reach 50% confluence, then use epidermal induction condition medium: low sugar DMEM / DF12 (1:1) medium (Sigma company, product number D-0547), 15ng / ml EGF (R&D company , product number 236-EG), 15ng / ml bFGF (R&D company, product number 234-FSE), 5ng / ml PDGF (R&D company, product number 222-AB), 1% ITS (Sigma company, product number I-3146), 10mmol / ml L dexamethasone (Sigma Company, product number D-1756), 100 IU / ml penicillin, 100 IU / ml streptomycin were used for induction culture, and epidermal cells could be obtained after 10-14 days. The results showed that 5 days after stem cel...

Embodiment 3

[0037] Embodiment 3, in vitro induction of MSC differentiation to dermal cells

[0038] The bone marrow-derived MSCs were planted on a well plate pre-coated with Matrigel gel, and low-sugar DMEM medium (Sigma Company, product number D-5523) containing 10% fetal bovine serum (Biochrom Company, product number S0115) was added, at 37 °C, 5%CO 2 Cultivate in the incubator until the cells reach 50% confluence, then use the dermal induction conditioned medium: high-glucose DMEM (Sigma company, product number D-5648), 5% fetal bovine serum (Biochrom company, product number S0115), 15ng / mlTGF-β 1 (R&D company, article number 240-B), 15g / mlbFGF (R&D company, article number 234-FSE), 1%ITS (Sigma company, article number I-3146), 10mmol / L dexamethasone (Sigma company, article number D-1756 ), 100IU / ml penicillin, and 100IU / ml streptomycin for induction culture, and dermal cells can be obtained after 10-14 days. The results showed that 10 days after stem cells were induced to differen...

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Abstract

The invention discloses a method to co-construct and organize engineering double layer skin in biological medicinal domain, which is characterized by the following: directional-evoking and differentiating stem cell of various tissue from outer to surface cell and dermal cells; getting the cell group; combining with biological material; constructing the method; preparing substitution good to restore and heal clinical burn, ulcer, would, deformed, defection after operation and scar on support stock. This method possesses larger market value latent force and wide prospect.

Description

technical field [0001] The present invention relates to the field of biomedicine, in particular to inducing differentiation of stem cells from different tissues to epidermis and dermis in vitro, and combining the induced differentiation cell groups with biological materials to jointly construct tissue engineering dual A method of layering skin, using this method to prepare skin tissue substitutes with specific shape and thickness for clinical burns, ulcers, wounds, deformities, postoperative defects, scars and other skin tissue repair and healing. Background technique [0002] As the largest organ of the human body, the skin has the functions of barrier, protection, temperature regulation and sensation. Skin defects caused by factors such as inflammation, ulcers, scalds, and burns can be life-threatening in severe cases. At present, autologous skin flaps or skin grafts are usually used to treat defect wounds in clinical practice, but they are faced with new wound defects in...

Claims

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Application Information

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IPC IPC(8): A61L27/60A61L27/38
Inventor 裴雪涛何丽娟陈琳南雪王韫芳管利东刘大庆白慈贤
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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