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Tissue engineered cornea epithelial transplantation membrane and preparation method and use thereof

A tissue engineering, corneal epithelial cell technology, applied in the fields of bioengineering and clinical medicine, to avoid immune rejection, good histocompatibility, and eliminate the effect of original antigenicity

Inactive Publication Date: 2012-01-18
PEKING UNIV THIRD HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no effective in vitro culture method for corneal endothelial cells, especially for primate corneal endothelial cells, especially for human corneal endothelial cells

Method used

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  • Tissue engineered cornea epithelial transplantation membrane and preparation method and use thereof
  • Tissue engineered cornea epithelial transplantation membrane and preparation method and use thereof
  • Tissue engineered cornea epithelial transplantation membrane and preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Embodiment 1, limbal tissue culture of human corneal epithelial cells in vitro

[0055] 1. In vitro primary and subculture corneal epithelial cells from corneal limbal tissue blocks

[0056] Primary culture: Disinfect human conjunctival sac and ocular surface with 10% povidone iodine, retrobulbar anesthesia with 2% lidocaine, use cataract phacoemulsification incision knife (Alcon, U.S.A.) to take a size of about 2.0mmX2.0mm in human corneal limbus The limbal tissue block with a thickness of about 200 μm was washed 3 times with D-PBS buffer containing 1% penicillin-streptomycin, then placed in 1.2U / ml dispase II digestion solution and kept overnight in the refrigerator at 4°C, after taking out the D- Wash with PBS buffer three times, and then treat with 0.25% trypsin-EDTA at 37°C for 10 minutes, and neutralize trypsin in DMEM medium containing 10% FBS. Place the limbal tissue pieces in a 60mm petri dish, dry naturally at room temperature in an ultra-clean bench, and adh...

Embodiment 2

[0077] Embodiment 2. Tissue engineered corneal stent material

[0078] 1. Carrier selection:

[0079] There are currently many kinds of carriers for constructing tissue-engineered corneal epithelial transplantation membranes, among which amniotic membranes, collagen membranes and hydrophilic gel membranes are mostly used, but these membranes all have a common disadvantage, that is, they lack a certain thickness to resist tension, compression and refraction. Therefore, its application is limited. Heterogeneous acellular matrix, especially porcine acellular matrix, is easy to obtain and has no antigenicity after enzymatic digestion of porcine corneal stromal cells, and has the structural characteristics that cornea should have.

[0080] 2. Preparation of porcine acellular corneal stroma:

[0081] Such as Figure 14 As shown, fresh pig eyeballs were soaked in 0.125% chloramphenicol solution for 1 hour, soaked in 75% ethanol for 0.15 hours, scraped off the epithelium with a ra...

Embodiment 3

[0082] Example 3: Preparation of Tissue-Engineered Corneal Epithelial Graft Membrane in Vitro

[0083] 1. Preparation of acellular porcine corneal stroma:

[0084] The corneal stroma prepared in Example 2 was soaked in fetal bovine serum for 24 hours and then placed in DMEM culture medium for future use.

[0085] 2. The corneal epithelial cells were subcultured in vitro and digested with 0.25% trypsin to make cell suspension.

[0086] 3. Place the decellularized porcine corneal stromal epithelium face up and place it in a culture dish to dry naturally for several minutes. 3 / mm 2 Inoculate on the decellularized porcine corneal stroma and wait for the cells to adhere to the wall in an ultra-clean bench, during which the culture solution is continuously dripped to avoid excessive drying of the cells. After the completion of wall attachment, a small amount of fetal bovine serum was added and placed in a 5% carbon dioxide incubator at 37°C for 24 hours. After 24 hours, an appro...

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Abstract

The invention discloses a tissue engineering cornea epithelial transplantation membrane, a preparing method and an application thereof. The invention aims to provide a tissue engineering cornea epithelial transplantation membrane, the tissue engineering cornea epithelial transplantation membrane comprises a porcine cornea epithelial cell and nonantigenic tissue engineering cornea bracket material, wherein, the porcine cornea epithelial cell and the nonantigenic tissue engineering cornea bracket material are combined into a whole by adopting a tissue engineering method. And meanwhile, The invention further provides a method for preparing the tissue engineering cornea epithelial transplantation membrane, and a use of the tissue engineering cornea epithelial transplantation membrane during the process of preparing medical treatment material used for remedying blind eye disease caused by the pathological state and the damage of corneas.

Description

technical field [0001] The invention relates to a tissue-engineered corneal epithelial transplantation membrane, its preparation method and application, and a scaffold material for constructing tissue-engineered cornea. The invention belongs to the fields of bioengineering and clinical medicine. Background technique [0002] Severe corneal lesions and blind eye diseases caused by corneal damage rank first among blinding eye diseases. Although corneal transplantation has a good therapeutic effect and a high success rate, the sources of donor corneal materials are very scarce. For a long time, it is urgently hoped that by recreating tissue-engineered biological cornea in vitro, the tissue-engineered cornea can become the equivalent of human cornea transplantation, and finally the successful transplantation of human cornea can restore the visual function of corneal blind patients and improve their quality of life . Therefore, seeking tissue-engineered corneas that can be use...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/36A61F2/14
Inventor 许永根王薇
Owner PEKING UNIV THIRD HOSPITAL
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