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Skin tissue engineering seed cells, construction method, adenovirus vector and use thereof

A technology of skin tissue engineering and seed cells, which is applied in the field of biomedicine, can solve problems such as immune rejection, and achieve the effect of reducing immunogenicity and immunogenicity

Inactive Publication Date: 2010-01-06
CHONGQING ZONGSHEN JUNHUI BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The purpose of the present invention is to provide a low immunogenic skin tissue engineering seed cell and its construction method, which can effectively solve the defect of immune rejection after transplantation of the existing allogeneic seed cells

Method used

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  • Skin tissue engineering seed cells, construction method, adenovirus vector and use thereof
  • Skin tissue engineering seed cells, construction method, adenovirus vector and use thereof
  • Skin tissue engineering seed cells, construction method, adenovirus vector and use thereof

Examples

Experimental program
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Effect test

no. 1 example

[0115] Step 1001, move the human skin taken from the circumcision into the ultra-clean operating table, rinse it with double-antibody PBS (containing 100u / ml penicillin and 100u / ml streptomycin; pH is 7.2) solution for 5 times, and put it in a petri dish Trim the subcutaneous fascia cells and excess subcutaneous fat, cut the trimmed human skin specimens into 0.3cm×2cm leather strips, then immerse them in Dispase II enzyme with a concentration of 2.5g / L, and digest them at 4°C for 12 hours . Remove the epidermis from the human skin specimen digested by Dispase II enzyme, cut the separated dermis into a paste with ophthalmic scissors, add collagenase at a concentration of 200U / ml and digest it in an incubator at a temperature of 37°C for 1 hour, and the tissue is basically The digestion was spread out to prepare a single cell suspension to the extent that no tissue pieces could be seen. Transfer the digest to a 10ml centrifuge tube, centrifuge and wash with PBS at 1500 rpm for ...

no. 2 example

[0166] Step 2001, move the human skin taken from the circumcision operation into the ultra-clean operating table, rinse 5 times with a double-antibody PBS (containing 100u / ml penicillin and 100u / ml streptomycin; pH is 7.2), and put it in a petri dish Trim the subcutaneous fascia cells and excess subcutaneous fat, cut the trimmed human skin specimens into 0.3cm×2cm leather strips, then immerse them in Dispase II enzyme with a concentration of 2.5g / L, and digest them at 4°C for 14 hours . Remove the epidermis from the human skin specimen digested by Dispase II enzyme, cut the separated dermis into a paste with ophthalmic scissors, add collagenase at a concentration of 200U / ml and digest it in an incubator at a temperature of 37°C for 2 hours, and the tissue is basically The digestion was spread out to the extent that no tissue pieces were visible, and a single cell suspension was prepared. Transfer the digest to a 10ml centrifuge tube, centrifuge and wash with PBS at 1500 rpm f...

no. 3 example

[0217] Step 3001, move the human skin taken from the circumcision into the ultra-clean operating table, rinse it with double-antibody PBS (containing 100u / ml penicillin and 100u / ml streptomycin; pH is 7.2) solution for 5 times, and put it in a petri dish Trim the subcutaneous fascia cells and excess subcutaneous fat, cut the trimmed human skin specimens into 0.3cm×2cm leather strips, then immerse them in Dispase II enzyme with a concentration of 2.5g / L, and digest them at 4°C for 16 hours . Remove the epidermis from the human skin specimen digested by Dispase II enzyme, cut the separated dermis into a paste with ophthalmic scissors, add collagenase at a concentration of 200U / ml and digest it in an incubator at a temperature of 37°C for 4 hours, and the tissue is basically The digestion was spread out to the extent that no tissue pieces were visible, and a single cell suspension was prepared. Transfer the digest to a 10ml centrifuge tube, centrifuge and wash with PBS at 1500 r...

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Abstract

The invention relates to skin tissue engineering seed cells, a construction method, an adenovirus vector and a use thereof, and the construction method comprises the steps of culturing human skin fibroblasts and constructing the adenovirus vector containing double genes of CTLA4Ig-CD40Ig; and the in vitro transfection of the human skin fibroblasts is carried out by the adenovirus vector containing the double genes of CTLA4Ig-CD40Ig. The skin tissue engineering seed cells with low immunogenicity are produced by the above construction method. The adenovirus vector used for transfection of the human skin fibroblasts is the Ad5F35 chimeric adenovirus vector which contains CTLA4Ig gene and CD40Ig gene effectively connected with a promoter. The invention relates to the use of adopting the adenovirus vector in the preparation of the transfected human skin fibroblasts for repairing injured skin. The invention can effectively reduce the immunogenicity of the seed cells, and the transfected human skin fibroblasts do not affect the proliferation performance thereof in vitro.

Description

technical field [0001] The invention relates to the field of biomedical technology, in particular to low-immunogenic skin tissue engineering seed cells and construction methods, adenovirus vectors and applications used in the process of skin injury and wound repair. Background technique [0002] Skin is an important tissue organ that covers and protects the body surface. Skin damage caused by various reasons needs to be repaired in time. For the repair of such defects, autologous skin grafting is often used, but this method not only causes new tissue defects in the donor area, but is also often limited by the source of the donor area. [0003] In recent years, the use of tissue engineering technology to produce tissue-engineered skin has become a hot spot in the treatment of skin defect transplantation. Tissue-engineered skin is generally inoculated with seed cells on scaffold materials in vitro, cultured in vitro to form a skin-like structure containing living cells, and ...

Claims

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Application Information

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IPC IPC(8): C12N15/62C12N15/861C12N5/10
Inventor 魏泓葛良鹏孔勇黄正根吴军
Owner CHONGQING ZONGSHEN JUNHUI BIOTECH
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