Novel decellularized amnion preparation method

A technology of decellularized amniotic membrane, applied in the field of preparation of decellularized amniotic membrane, which can solve the problems of remaining extracellular matrix composition, biological activity and biomechanical properties, etc., avoiding the use of trypsin chemical reagents, reducing residues, retaining The effect of completeness

Inactive Publication Date: 2019-03-01
广州瑞铂茵健康科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Although chemical treatment is capable of removing cellular components, chemical treatment also has some adver

Method used

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  • Novel decellularized amnion preparation method
  • Novel decellularized amnion preparation method
  • Novel decellularized amnion preparation method

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preparation example Construction

[0056] The invention provides a method for preparing the decellularized amniotic membrane. The preparation method comprises ultrasonically treating the amniotic membrane, and then physically scraping off the cells on the amniotic membrane to obtain the decellularized amniotic membrane. The preparation method of the acellular amnion provided by the present invention adopts ultrasonic treatment and physical scraping to remove the cells on the surface of the amnion, avoids the use of trypsin chemical reagents, and the prepared acellular amnion product is safer and reduces the harmful effects of the amnion on the body. Residues of harmful chemicals, while retaining the integrity of the amniotic matrix to the greatest extent.

[0057] In some preferred embodiments, the preparation method further includes removing residual blood in the amniotic membrane before sonication. Optionally, use buffers such as phosphate buffer or carbonate buffer that can maintain the osmotic pressure and ...

Embodiment 1

[0079] This embodiment provides a method for preparing decellularized amniotic membrane, the method comprising the following steps:

[0080] (a) After washing the blood residue in the amniotic membrane with physiological saline, soak the amniotic membrane with water for 12 hours;

[0081] (b) spread the amniotic membrane on the bottom of the petri dish, seal the petri dish and place it in an ultrasonic cleaner for 2 hours of sonication, the ultrasonic power is 600W, and the ultrasonic frequency is 40kHz;

[0082] (c) Gently scrape off the cells remaining on the surface of the amniotic membrane with a cell scraper;

[0083] (d) Steps (b) to (c) are repeated once, and then the prepared decellularized amniotic membrane is spread on the surface of a clean culture dish to dry.

Embodiment 2

[0085] This embodiment provides a method for preparing decellularized amniotic membrane, the method comprising the following steps:

[0086] (a) After washing the blood residue in the amniotic membrane with physiological saline, soak the amniotic membrane with water for 6 hours;

[0087] (b) spread the amniotic membrane on the bottom of the petri dish, seal the petri dish and place it in an ultrasonic cleaner for 4 hours of sonication, the ultrasonic power is 500W, and the ultrasonic frequency is 50kHz;

[0088] (c) Gently scrape off the cells remaining on the surface of the amniotic membrane with a cell scraper;

[0089] (d) Steps (b) to (c) are repeated once, and then the prepared decellularized amniotic membrane is placed on the surface of a clean culture dish and spread to dry.

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Abstract

The invention provides a novel decellularized amnion preparation method, and relates to the technical field of biological materials. The method comprises ultrasonically treating the amnion and then physically scraping cells on the amnion to obtain the decellularized amnion. According to the method, the ultrasonic treatment and physical scraping mode is adopted to remove the cells on the surface ofthe decellularized amnion, the use of chemical reagents like trypsin is avoided, and the prepared decellularized amnion product is safer and reduces the residual chemical substances harmful to the body in the decellularized amnion. At the same time, the integrity of the amniotic matrix is retained to the utmost extent.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a novel preparation method of decellularized amniotic membrane. Background technique [0002] Scaffold materials used in tissue engineering refer to materials that can support specific cells, guide tissue regeneration and control tissue structure, and can be implanted into organisms and combined with tissue living cells. The scaffold material in tissue engineering plays an extremely important role in tissue engineering. It can regulate the growth and biological functions of cells in the scaffold, and at the same time promote the differentiation of cells loaded in the scaffold to the target tissue or organ. Provides initial support for cells, positions cells in a suitable space, provides physical and biological cues for cell adhesion, migration, proliferation and differentiation, and propagates cells and assembles their secreted extracellular matrix into functional ti...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61L27/50
CPCA61L27/3633A61L27/3691A61L27/50A61L2430/40
Inventor 嵐山芮魏伟许超
Owner 广州瑞铂茵健康科技有限公司
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