Ovarian extracellular matrix scaffold and hydrogel as well as preparation method and application thereof

A technology of ovarian cells and extracellular matrix, which is applied in the fields of pharmaceutical formulation, medical science, tissue regeneration, etc., and can solve the problems of limited research on decellularization

Active Publication Date: 2021-04-30
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] However, decellularization research in the field of female reproduction is currently very limited

Method used

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  • Ovarian extracellular matrix scaffold and hydrogel as well as preparation method and application thereof
  • Ovarian extracellular matrix scaffold and hydrogel as well as preparation method and application thereof
  • Ovarian extracellular matrix scaffold and hydrogel as well as preparation method and application thereof

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preparation example Construction

[0070] The invention discloses a method for preparing an ovarian extracellular matrix scaffold, which is characterized in that it comprises the following steps:

[0071] 1) Pretreatment of ovarian tissue: remove the fatty tissue around the mammalian ovary, separate the ovarian tissue skin and medulla, and release the follicular fluid;

[0072] 2) Multiple decellularization treatments: Take the ovarian tissue skin and medulla treated in step 1) and place them in double distilled water, sodium deoxycholate and trinaton X-100 mixed solution, lauryl sulfate Shake and wash in sodium solution, then add DNase and incubate in medium at 37°C for 4-8 hours;

[0073] 3) Sterilization and freeze-drying treatment: take the intermediate product after step 2) and place it in a disinfectant solution for 4-6 hours, and then place it in a vacuum environment for 16-24 hours and freeze-dry it to obtain a freeze-dried tissue;

[0074] 4) Preparation of ovarian extracellular matrix scaffold: take ...

Embodiment 1

[0092] This embodiment discloses a method for preparing ovarian extracellular matrix hydrogel containing granulosa cells and follicles, which includes the following preparation steps:

[0093] 1) Pretreatment of ovarian tissue: obtain the pig ovarian tissue of the animal from the slaughterhouse, separate the skin and medulla of the pig ovarian tissue, and release the follicular fluid; the specific steps include: the pig ovarian tissue is placed in a phosphate-buffered saline In the solution container, the whole porcine ovary was placed in a storage box containing dry ice and transported to the laboratory. Subsequent experiments were performed in an ultra-clean workbench. The ultra-clean workbench was wiped with alcohol and irradiated with ultraviolet light 1 hour in advance. Use tissue scissors to remove the fat tissue around the porcine ovary, cut the porcine ovary from the hilum in a butterfly shape, separate the skin and medulla, control the thickness of the ovarian cortex t...

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Abstract

The invention discloses an ovarian extracellular matrix scaffold and hydrogel as well as a preparation method and application thereof, and belongs to the technical field of biomedicine and clinical medicine. The preparation method of the ovarian extracellular matrix scaffold comprises the following steps: 1) ovarian tissue pretreatment; 2) multi-decellularization treatment: sequentially putting the ovarian tissue skin and marrow treated in the step 1) into double distilled water, a mixed solution of sodium deoxycholate and triton X-100 and a lauryl sodium sulfate solution, sufficiently shaking and washing, adding DNA enzyme, and culturing in a culture medium at 37 DEG C for 4-8 hours; (3) sterilization and freeze-drying treatment: (4) preparation of the ovarian extracellular matrix scaffold: grinding the freeze-drying tissue prepared in the step (3) into particles with the particle size of less than 1mm, and adding acid liquor, pepsase, alkali liquor and a buffer solution into the particles to prepare the ovarian extracellular matrix scaffold. and continuously adding granular cells and follicles into the ovarian extracellular matrix scaffold to prepare the hydrogel material which can be implanted into a mammal body and has the effects of repairing ovarian injury and reconstructing ovarian functions. The research provides a new strategy for female fertility protection.

Description

technical field [0001] The invention relates to a material for protecting ovarian function, which belongs to the technical field of clinical medical treatment, and in particular relates to an ovarian extracellular matrix scaffold and hydrogel, a preparation method and application thereof. Background technique [0002] Factors such as premature ovarian insufficiency, autoimmune diseases, and tumor treatment may destroy ovarian function. In addition, premature ovarian failure caused by multiple factors such as genetics, environment, social psychology, and lifestyle will cause female physical and mental health. Mental health problems. [0003] However, women with compromised ovarian function still have other options to restore fertility. At present, there are more than 200 babies born through ovarian transplantation in the world. Although the effectiveness of ovarian transplantation has been confirmed more and more, for women who are treated for primary tumors, there may be re...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/50A61L27/52
CPCA61L27/3633A61L27/3641A61L27/3687A61L27/3691A61L27/50A61L27/52A61L2430/40A61L2300/412A61L2400/06
Inventor 王世宣张金金吴桐
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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