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Preparation method of tissue engineering cornea

A tissue engineering and corneal technology, applied in the field of tissue engineering cornea and its preparation, can solve the problems of limited source of corneal limbal stem cells, poor clinical application effect, inability to repair corneal defects, etc. The effect of large immune rejection and low cost

Active Publication Date: 2013-04-24
SHAANXI RUISHENG BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The current research on tissue engineering cornea mainly focuses on using the amniotic membrane that removes cell components as a carrier, and inoculates limbal stem cells on its surface to construct. Repair corneal defects, making it less effective 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 stents: Obtain bovine corneal tissue, peel off the pericorneal tissue, wash with PBS solution and freeze at -80°C for 1 hour, take it out and thaw at room temperature, and repeat freezing and thawing 4 times to make the cells Completely rupture and 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% protease solution for 3 hours, rinse with pure water for 3 to 5 times; put it in Soak in 0.8M NaOH solution for 20 minutes to achieve the purpose of lysing cells and inactivating viruses, rinse with PBS solution to neutral pH; Soak in the mixed solution for 30 minutes to remove residual DNA and α-galactosyl (α-gal) antigen components, reduce immunogenicity, rinse with PBS solution; use 8% (V / V) RGD polypeptide solution after dehydration and dryin...

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Abstract

The invention relates to a preparation method of a tissue engineering cornea, comprising the following steps of: by using epidermal stem cells and pluripotent stromal stem cells as seed cells, planting on the two surfaces of an acellular natural cornea stroma after in-vitro amplification culture; and then forming the tissue engineering cornea through in-vitro induction culture. A scaffold used inthe preparation method not only overcomes the problem of serious immunological rejection of an xenogeneic cornea stroma, but also retains the structure (capable of recovering the transparency of the cornea) and main components (comprising a growth factor capable of promoting the growth, the proliferation and the differentiation of cornea cells) of a natural cornea. Compared with a product of the prior art, the invention has low cost, simple and convenient operation, extensive sources and easy storage; and in addition, the tissue engineering cornea containing living cells has certain elasticity and toughness, easy changes on shape, size and thickness and extremely low immunogenicity, can prevent complications caused by non-corneal materials and be rebuilt by recipient cells when implanted in vivo and rapidly integrated with an organism to realize transparency and is suitable for repairing corneal injury caused by various reasons.

Description

technical field [0001] The invention belongs to the technical field of tissue engineering of biological materials, and in particular relates to a tissue engineering cornea and a preparation method thereof. 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 other 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 transplantation is the most effective means to treat corneal lesions, and the donors of traditional corneal transplantation mainly come from cadavers and donations. At present, there are about 4 million patients with corneal blindness in my country, and many of them can be cured by corneal transplantation. H...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/36
Inventor 王爱军
Owner SHAANXI RUISHENG BIOTECH
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