Preparation method and application of composite decellularized amniotic membrane of endometrial stem cells

A technology of endometrial stem cells and decellularization, which is applied in the field of preparation of endometrial stem cells combined with decellularized amnion, can solve the problems of endocrine interference in patients, achieve good repair effect and avoid interference

Inactive Publication Date: 2020-04-24
杭州恩格生物医疗科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the technical problem existing in the prior art that repairing the endometrium with hormones easily interferes with the endocrine of p

Method used

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  • Preparation method and application of composite decellularized amniotic membrane of endometrial stem cells

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Experimental program
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Embodiment 1

[0032] S1) The fresh full-term placenta was repeatedly washed with normal saline until there was no obvious blood on the surface, and then the amnion and chorion were peeled off to leave the complete amnion tissue including the epithelial cell layer, basement membrane and avascular matrix, and then the mass fraction was Rinse the intact amnion tissue once with 3% hydrogen peroxide solution, and then rinse twice with PBS solution with a pH of 7.2-7.4;

[0033] S2) Treat the intact amniotic membrane tissue treated by S1) with 0.2% EDTA at 35°C for 30 minutes, then soak it in 0.3mol / L sodium hydroxide solution for 20s, and then transfer it to 3% concentration Shake vigorously in ammonium chloride to remove cells and make translucent amniotic membrane;

[0034] S3) Wash the translucent amniotic membrane twice with PBS solution with a pH of 7.2-7.4, and then immerse and sterilize it in physiological saline containing antibiotics for 10 minutes; wherein, the antibiotics contained in...

Embodiment 2

[0040] S1) The fresh full-term placenta was repeatedly washed with normal saline until there was no obvious blood on the surface, and then the amnion and chorion were peeled off to leave the complete amnion tissue including the epithelial cell layer, basement membrane and avascular matrix, and then the mass fraction was Rinse the intact amnion tissue twice with 4% hydrogen peroxide solution, and then rinse three times with PBS solution with a pH of 7.2-7.4;

[0041] S2) Treat the intact amniotic tissue treated by S1) with 0.4% EDTA at 36°C for 40 minutes, then soak in 0.4mol / L sodium hydroxide solution for 25s, and then transfer to 4% concentration Shake vigorously in ammonium chloride to remove cells and make translucent amniotic membrane;

[0042] S3) The translucent amniotic membrane was washed three times with PBS solution with a pH of 7.2-7.4, and then soaked and disinfected in physiological saline containing antibiotics for 13 minutes; wherein, the antibiotics contained ...

Embodiment 3

[0048] S1) The fresh full-term placenta was repeatedly washed with normal saline until there was no obvious blood on the surface, and then the amnion and chorion were peeled off to leave the complete amnion tissue including the epithelial cell layer, basement membrane and avascular matrix, and then the mass fraction was Rinse the intact amnion tissue twice with 5% hydrogen peroxide solution, and then rinse four times with PBS solution with a pH of 7.2-7.4;

[0049] S2) Treat the intact amniotic tissue treated by S1) with 0.5% EDTA at 37°C for 45 minutes, then soak in 0.5mol / L sodium hydroxide solution for 30s, and then transfer to 5% concentration Shake vigorously in ammonium chloride to remove cells and make translucent amniotic membrane;

[0050] S3) The translucent amniotic membrane was washed 4 times with PBS solution with a pH of 7.2-7.4, and then soaked and disinfected in physiological saline containing antibiotics for 15 minutes; wherein, the antibiotics contained in th...

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Abstract

The invention discloses a preparation method and application of a composite decellularized amniotic membrane of endometrial stem cells in the technical field related to physiological health care, including preparation of complete amniotic membrane tissue, preparation of a translucent amniotic membrane, immersion disinfection, decellularization treatment, preparation of a stem cell amniotic membrane culture system, preparation and cultivation of endometrial stem cell composite decellularized amniotic membrane and other processes as well as cryopreservation, thawing and application. The endometrial stem cell composite decellularized amniotic membrane prepared by the method is applied to the preparation of endometrial repair materials, is transplanted into the endometrial injury site for repair of endometrial damage and increase the thickness of the endometrium; the decellularized amniotic membrane completely removes cells and nucleic acids, and does not produce rejection reactions in thebody; through combination of menstrual blood stem cells or endometrium stem cells with the decellularized amniotic membrane, the cells are homologous to endometrial cells, and the repair effect is good; the use of hormones in repair is completely excluded to avoid interference with the internal secretion of a human body.

Description

technical field [0001] The invention relates to the technical field related to physiological health care, in particular to a preparation method and application of endometrial stem cells compounded with acellular amnion. Background technique [0002] The endometrial layer is the part that constitutes the inner wall of the mammalian uterus. It is divided into two layers: the functional layer and the basal layer. Two-thirds of the endometrial surface is the dense layer and the spongy layer, collectively referred to as the functional layer, which responds to both estrogen and progesterone; The basal layer is the 1 / 3 endometrium close to the myometrium, which is not affected by ovarian sex hormones and does not undergo periodic changes. [0003] The thickness of the endometrium changes at different times such as ovulation and menstruation, that is, it gradually becomes thicker from very thin, and then sheds during menstruation, and so on. Thin endometrium is a common disease in ...

Claims

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Application Information

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IPC IPC(8): A61L27/38A61L27/36A61L27/50
CPCA61L27/3604A61L27/3687A61L27/3691A61L27/3834A61L27/50A61L2430/22A61L2430/40
Inventor 王健侯雷何灿洋何灿平竺亚斌
Owner 杭州恩格生物医疗科技有限公司
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