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Method for preparing dried amnion

An amniotic membrane drying technology, applied in medical science, prostheses, etc., can solve the problems that the effect has not been confirmed, and achieve the effect of complete cell phenotype and basement membrane, easy implementation, and optimized amniotic membrane characteristics

Inactive Publication Date: 2011-07-27
周海华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The amniotic membrane preserved by this method is closer to the natural fresh amniotic membrane in terms of morphology and biology, but the effect has not been further confirmed

Method used

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  • Method for preparing dried amnion
  • Method for preparing dried amnion
  • Method for preparing dried amnion

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] The preparation method of the present invention comprises the following steps: the donor is taken from the fetal membrane obtained by cesarean section, and the preoperative virological test results of the puerpera are all negative; Bacterial saline was used to wash the blood on the surface; the operator wore sterile gloves on both hands, gently separated the amniotic membrane from the chorionic membrane, rinsed it with sterile BSS solution, spread it on a sterile glass plate, and scraped it with a cell scraper. Remove the spongy layer, fibroblast layer and serous exudate; soak and disinfect the fresh amniotic membrane with PBS containing 50,000 U / L penicillin, 80,000 U / L tobramycin, and 100 mg / L streptomycin for 3 Time × 5min; at -20°C-4°C, soak the sterile amniotic membrane in ECM glue for 15min-30min; drop 0.25% sterile sodium fluorescein solution 0.2ml in the fume hood at 16°C-37°C Stain the amniotic membrane for 5 minutes, then lay the dyed amniotic membrane on the ...

Embodiment 2

[0040] The preparation method of the present invention comprises the following steps: the donor is taken from the fetal membrane obtained by cesarean section, and the preoperative virological test results of the puerpera are all negative; Bacterial saline was used to wash the blood on the surface; the operator wore sterile gloves on both hands, gently separated the amniotic membrane from the chorionic membrane, rinsed it with sterile BSS solution, spread it on a sterile glass plate, and scraped it with a cell scraper. Remove the spongy layer, fibroblast layer and serous exudate; soak and disinfect the fresh amniotic membrane with PBS containing 50,000 U / L penicillin, 80,000 U / L tobramycin, and 100 mg / L streptomycin for 3 Time × 5min; at -20°C-4°C, soak the sterile amniotic membrane in FN glue for 15min-30min; drop 0.1% sterile trypan blue solution 0.2ml in the fume hood at 16°C-37°C Stain the amniotic membrane for 5 minutes, then lay the dyed amniotic membrane on the nitrocell...

Embodiment 3

[0042] The preparation method of the present invention comprises the following steps: the donor is taken from the fetal membrane obtained by cesarean section, and the preoperative virological test results of the puerpera are all negative; Bacterial saline was used to wash the blood on the surface; the operator wore sterile gloves on both hands, gently separated the amniotic membrane from the chorionic membrane, rinsed it with sterile BSS solution, spread it on a sterile glass plate, and scraped it with a cell scraper. In addition to its spongy layer, fibroblast layer and serous exudate. Soak fresh amnion in PBS containing 50,000 U / L penicillin, 80,000 U / L tobramycin, and 100 mg / L streptomycin for 3 times×5min; Soak the amniotic membrane in Matrige gel for 15min-30min; in a fume hood at 16°C-37°C, drop 0.2ml of 0.5% sterile tiger red solution on the amnion and stain it for 5min, then spread the stained amnion on the nitrocellulose membrane , in a sterile fume hood to dry natur...

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Abstract

The invention relates to a method for preparing dried amnion, which is characterized by comprising the following steps of: strictly sterilizing fresh amnion which carries no virus and is repeatedly rinsed, rebuilding and repairing ground substances at low temperature, unfolding the amnion, drying, sterilizing by cobalt 60 and preserving to reserve. The method for preparing the dried amnion has the advantages that the manufacturing process is simple and reliable and the cost is relatively low to ensure that the implementation and the popularization are easy; the prepared dried amnion can be preserved at normal temperature and conveniently transported; the cell phenotype and the basement membrane of the amnion are intact; fewer amnion ground substances are destroyed; the prepared dried amnion is superior to purely lyophilized amnion preserved at the same time and the like.

Description

technical field [0001] The invention relates to a method for preparing amniotic membrane, in particular to a method for preparing dried amniotic membrane. Background technique [0002] The amniotic membrane is derived from the embryo and is a transparent, nerveless, vascular and lymphatic, low antigenic and resilient tissue. The membrane is composed of epithelial cell layer, basement membrane, stroma layer, fibroblast layer and spongy layer, with a thickness of about 0.02-0.05 mm. The amniotic epithelium is a single-layer cuboidal epithelium, with many microvilli on the surface and sides of the cells, which have the ability to synthesize, secrete and deposit basement membrane and extracellular matrix components, have the potential of differentiation, and can secrete a variety of growth factors; while the amnion The matrix is ​​rich in collagen fibers and various forms of protease inhibitors; the amnion basement membrane is the thickest basement membrane in the human body (a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36
Inventor 周海华
Owner 周海华
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