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Active gene modified pig skin for covering and repairing human skin wounds and application of active gene modified pig skin

A technology for skin wounds and genetic modification, applied in skin transplantation, gene therapy, genetic engineering, etc., can solve problems such as loss of function, achieve the effect of promoting wound repair, solving the problem of inability to survive, and saving the lives of patients

Active Publication Date: 2021-06-22
GENEO MEDICINE CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, wild-type pigs will soon (within 3-4 days) be rejected by hyperacute immune rejection after the full-thickness skin coverage restores blood supply, and will soon lose their function

Method used

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  • Active gene modified pig skin for covering and repairing human skin wounds and application of active gene modified pig skin
  • Active gene modified pig skin for covering and repairing human skin wounds and application of active gene modified pig skin
  • Active gene modified pig skin for covering and repairing human skin wounds and application of active gene modified pig skin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Antigen removal from isolated porcine skin and human protein expression and functional identification, Figure 2-5 The flow cytometry test indicated that the genetically modified pigs stably expressed human-derived CD55, CD59, CD46, CD47, CD39, THBD, TFPI and HLAE proteins, indicating that the above genes were successfully knocked in. The expression of GGTA, B4gal, and Camh proteins showed that the galactose antigen gene knockout was successful.

[0047] Figure 6-8 Antibody binding experiment of the mixed human serum shown: Pig ear fibroblasts were incubated with 30 healthy volunteers mixed serum at 37°C for 30 minutes with 75% (100% human mixed serum normal saline mass fraction diluted to 75% before the experiment), Wild-type pig cells carry galactose antigens that can be bound by pre-stored antibodies in human serum. Fluorescent antibody staining marks human IgM and IgG antibodies bound to pig cells, and the fluorescence intensity is measured by flow cytometry.

[...

Embodiment 2

[0051] Ex vivo porcine skin-Stone crab monkey skin xenograft

[0052] 1. Pig skin preparation: Inject phenobarbital sodium at a dose of 30 mg / Kg for anesthesia. After shaving, scrub repeatedly with soap and tap water, then routinely disinfect with povidone iodine and 70% alcohol by volume, spread towels, and take the back The skin is made of pigskin slices with a thickness of about 0.6mm using an electric skin remover, cut into 5cm×5cm skin slices, rinsed repeatedly with physiological saline containing antibiotics and placed in a 4-degree refrigerator for later use. In addition, fresh skin slices were infiltrated in an antifreeze protective agent containing 5% DMSO for 20 minutes, then taken out, packed into a sterile polyvinyl chloride pouch, sealed, and placed in a -80°C refrigerator or liquid nitrogen tank for later use. Before use, rewarm in a 37°C water bath and rinse with normal saline 3 times.

[0053] 2. Transplantation steps of genetically modified pig full-thickness...

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Abstract

The invention provides active gene modified pig skin for covering and repairing human skin wounds and an application of the active gene modified pig skin, pig immune antigens GGTA, B4GAL and CAMH are removed from the pig skin, and human CD55, 59, 46, 47, 39, 141, HLEA and TFPI protein combinations are carried. The obtained in-vitro pig skin can be used for covering and sealing a skin wound surface, can recover effective blood circulation and complete skin functions of the wound surface, can save the life of a patient, can promote wound surface repair, can reduce scar formation, and can solve the problem that the pig skin cannot survive when the skin wound surface is transplanted in the prior art.

Description

technical field [0001] The invention belongs to the technical field of biogenetic engineering, and relates to an active genetically modified pig skin for covering and repairing human skin wounds and an application thereof. Background technique [0002] The difficulty in the clinical management of trauma, especially large area burns, lies in the initial wound coverage. The current clinical strategy is early debridement and autologous skin grafting. However, large burn wounds often do not have enough autologous skin grafts to cover them, and must be temporarily covered with allogeneic human cadaver skin or artificially synthesized skin materials, but such skin sources are very limited. In addition, severe trauma with skin and soft tissue defects in critical condition often cannot tolerate the first-stage skin graft after early expansion, and the wound needs to be temporarily covered to delay skin grafting. Porcine skin coverings currently in clinical use are essentially inac...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/60C12N5/10C12N15/113C12N15/85C12N15/90
CPCA61L27/3804A61L27/60C12N15/113C12N15/85C12N15/907C07K14/47C07K14/70596C07K14/70503C07K14/745A61L2300/412C12N2310/20A61L27/38C12N5/10C12N15/90A61F2/10A61K48/00C12N15/09C07K14/435
Inventor 王刚
Owner GENEO MEDICINE CO LTD
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