Construction method and application for complexion adjustable organization engineering skin

A technology of tissue engineering and construction methods, which is applied in the direction of skin transplantation, artificial cell constructs, biochemical equipment and methods, etc., and can solve problems such as inability to repair skin grafts

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

[0008] The tissue-engineered skin currently used in clinical practice cannot be repaired according to the skin color of the patient.

Method used

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  • Construction method and application for complexion adjustable organization engineering skin
  • Construction method and application for complexion adjustable organization engineering skin
  • Construction method and application for complexion adjustable organization engineering skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] Example 1. Taking bone marrow MSC as an example, stem cells from different tissue sources were isolated in vitro

[0038] 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

[0039] Embodiment 2, in vitro induction of MSC differentiation to epidermal cells

[0040] The bone marrow-derived MSCs were planted on a well plate pre-coated with Matrigel gel (BD company, product number 354240), and low-sugar DMEM medium (Sigma company, product number D- 5523), at 37°C, in a 5% CO2 incubator, cultivate until the cells reach 50% confluence, then use the epidermis induction conditioned 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, article number I-3146), 10mmol / L dexamethasone (Sigma company, article number D-1756), 100IU / ml penicillin, 100IU / ml streptomycin for induction culture, and epidermal cells can be obtained after 10-14 days. The results showed that 5 days after stem cells were induced to differentiate into epidermal cells, the cells showed th...

Embodiment 3

[0041] Embodiment 3, in vitro induction of MSC differentiation to dermal cells

[0042] 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-β1 (R&D company, article number 240-B), 15g / ml bFGF (R&D company, article number 234-FSE), 1%ITS (Sigma company, article number I-3146), 10mmol / L dexamethasone (Sigma company, Product No. D-1756), 100IU / ml penicillin, 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 differentiate int...

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Abstract

The invention discloses a method to construct engineering skin to adjust flesh in biological medicine domain, which comprises the following steps: orient-evoking and differentiating stem cell of diverse tissue to sheath and dermal cell; combining the cell group and culturing melanin cell with biological material at a finite proportion; preparing substitution good to restore and heal clinical burn, ulcer, would, deformed, defection after operation and scar. This invention can meet diverse needs of individual flesh according to add diverse melanin cell, which possesses giant latent capacity and wide application prospect.

Description

technical field [0001] The 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 differentiated cell groups and isolated cultured melanocytes with biological A method of combining materials to jointly construct tissue-engineered skin that can adjust skin color. Using this method, it can be used for the repair and healing of clinical burns, ulcers, trauma, deformities, postoperative defects, scars and other skin tissues with specific shapes and thicknesses. , A skin tissue substitute that can adjust skin tone. 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 ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/08A61F2/10C12N5/071
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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