Preparation method of amnion tissue engineering de-immunized skin scaffold

Abstract

The invention discloses a preparation method of an amnion tissue engineering de-immunized skin scaffold, comprising the following steps: rinsing and disinfecting amnion, then immersing in a commercialamnion cell culture medium, culturing in a CO2 incubator at 37 DEG C for 72 h, taking the cultured amnion out of the culture medium, centrifuging the cultured amnion and filtering through a 0.22 micron filter membrane and then sub-packaging, and cryopreserving in a refrigerator at 80 DEG C; carrying out continuous gradient dehydration on the amnion with ethanol for 3 days, aseptically air-drying,cutting into different specifications of amnion patches according to clinical requirements, and carrying out aseptic vacuum package for later use. The method adopted by the invention can realize thepurpose of simple, rapid, high-efficiency and large-scale preparation of medical artificial skin; the material used is human or pig amnion, has a wide source, is mostly waste after production and is low-cost, and there is no ethical problem. The invention has been verified on skin regeneration after burns in rats and pigs (large animal models), and the effect is remarkable and no adverse immune reaction is observed.

Description

technical field [0001] The invention relates to a preparation method of amnion tissue engineering deimmunized skin scaffold. Background technique [0002] Amnion tissue discarded during childbirth is one of the important sources of seed cells for regenerative medicine. It has a wide range of sources, is easy to obtain, and has no ethical and moral disputes. It has become a research hotspot in recent years. Amnion, as an immune-exempt biological graft, has biological properties such as anti-inflammation, anti-adhesion, pain relief, and promotion of epithelialization. It has been applied to burn patients and other surgical operations as early as 1910 to promote epidermalization, Reduce pain and prevent infection. In recent years, with the further understanding of amniotic membrane and the development of amniotic membrane preservation and preparation technology, amniotic membrane has once again become a research and application hotspot, and is widely used in corneal reconstruc...

AI Technical Summary

Problems solved by technology

[0004] At present, amniotic membrane is used as a skin tissue repair material, and the commonly used technologies are fresh amniotic membrane or deep-frozen amniotic membrane, but there are still defects such as the introduction of immunogenicity by allogeneic cells and the inconvenience of storage, which cannot be taken at any time.

Follow-up series of decellularized amniotic membrane skin scaffolds came into being (such as CN105688287A, CN105797212A, CN106668955A, CN107019816A, CN107254431A, 101330935A, CN107233623A, CN1757717A, CN102327640A etc.), but these technologies mostly adopt methods such as digestion of pancreatic amniotic membrane cells The operation is cumbersome, and it is difficult to overcome the defects that the amniotic membrane is jelly-like after digestion with trypsin, contains impurities such as cell proteins, causes allergic reactions, and cannot cover wounds with damaged skin.

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