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Method for preparing tissue engineering cornea

A tissue engineering and cornea technology, applied in the field of tissue engineering cornea preparation, can solve the problems of the amniotic membrane that has been removed from the cell components cannot exert amniotic membrane stem cells, the source of corneal limbal stem cells is limited, and the clinical application effect is not good, and the ability to proliferate and differentiate Strong, easy to change shape size and thickness, low cost effect

Active Publication Date: 2013-04-24
SHAANXI RUISHENG BIOTECH
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Problems solved by technology

[0004] The current tissue-engineered cornea mainly uses amniotic membrane with decellularized components as a carrier, and limbal stem cells are inoculated on the surface to construct it. The main disadvantage is that the source of limbal stem cells is limited, and the amniotic membrane depleted of cellular components cannot play the role of amniotic membrane stem cells; at the same time The amniotic membrane structure is very thin, has no strength, and cannot repair corneal defects, making it ineffective in clinical application

Method used

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Embodiment Construction

[0014] The technical solutions of the present invention will be further described in detail below in conjunction with specific examples.

[0015] Step 1. Preparation of corneal stent: Obtain porcine corneal tissue, peel off the pericorneal tissue, wash with PBS solution and freeze at -80°C for 1 hour, take it out and thaw it at room temperature, and repeat freezing and thawing for 3 times to completely rupture the cells Disintegrate; soak in pure water at 4°C for 1 day, make it swell and cut off 1 / 2 thickness; then digest it in 0.2% (w / v) protease solution for 2 hours, rinse with pure water for 3 to 5 times ; Soak it in 0.5M NaOH solution for 20 minutes to achieve the purpose of lysing cells and inactivating viruses, rinse with PBS solution until the pH is neutral; Soak in the mixed solution of glycosidase for 30 minutes to remove residual DNA and α-galactosyl antigen components, reduce immunogenicity, rinse with PBS solution; use 10% (v / v) polylysine after dehydration and dry...

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Abstract

The present invention discloses a producing method for tissue engineering cornea. The method involves using amniotic epithelial cells and amniotic stromal cells as seed cells, planting at two sides of decellularized natural corneal stroma after inducement and differentiation in vitro, forming tissue engineering cornea through organ culture in vitro. The scaffold of present invention not only resolves the problem of strong immunological rejection of heterogenetic corneal stroma, but also sustains the structure (which can restore transparency of cornea) and major components (including growth factors which could promote corneal cell growth, proliferation and differentiation) of natural cornea. Compared with the prior art, present invention has advantages of low cost, easy operation, wide range of sources and easy preservation. The said tissue engineering cornea containing live cells has certain elasticity and tenacity, and its shape, size and thickness can be changed easily. The tissue engineering cornea has very low immunogenicity. It could avoid complications caused by non-corneal material, be reconstructed by recipient cells after being transplanted in body, and be incorporated into organism rapidly to become transparent. It could be used to repair corneal wounds caused by all kinds of reasons.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering of biological materials, and in particular relates to a preparation method of tissue engineering cornea. Background technique [0002] The anatomical characteristics of the eyeball determine that the position of the eyeball is exposed, the tissue structure is fragile, and a small external force or foreign body can cause serious damage. In addition, some corneal diseases include infectious keratopathy, corneal degeneration, malnutrition and immune keratopathy, etc. Can cause corneal damage. Once damaged, it will seriously affect vision or cause blindness. Corneal lesions are the second leading cause of blindness next to cataracts, and the number of cases is increasing at a rate of 1.5 million to 2 million cases per year. While keratoplasty is the most effective means of treating corneal lesions, donors for traditional keratoplasty mainly come from cadavers and donations. At present, t...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/36
CPCC12N2506/02C12N2533/90A61L2430/16A61F2/142C12N5/0605C12N5/0621
Inventor 王爱军
Owner SHAANXI RUISHENG BIOTECH
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