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Method for preparing amniotic mesenchymal stem cells from human placental amniotic membrane and its application

A technology of stem cells and human amniotic membrane, applied in the field of stem cell therapy, can solve problems affecting cell separation and culture, high contamination rate of human amniotic membrane collection, poor cell activity, etc.

Active Publication Date: 2019-09-20
BOYALIFE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The invention solves the problems in the prior art that there is a high contamination rate of human amniotic membrane collection and poor cell activity after collection, which directly affects subsequent cell separation and culture

Method used

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  • Method for preparing amniotic mesenchymal stem cells from human placental amniotic membrane and its application
  • Method for preparing amniotic mesenchymal stem cells from human placental amniotic membrane and its application
  • Method for preparing amniotic mesenchymal stem cells from human placental amniotic membrane and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0088] Example 1: Preparation of human amniotic mesenchymal stem cells from placental amniotic membrane

[0089] (1) Take out the placenta from the placenta sample collection box, place it in a white porcelain dish, and use a basic balanced salt solution (0.4g of KCl, 0.06g of KH 2 PO 4 , 0.132g of Na 2 HPO 4 .12H 2 O, 8g of NaCl, 0.35g of NaHCO 3 , the D-glucose of 1.0g, the streptomycin of 0.10g, the penicillin of 0.06g are dissolved in water and be settled into the solution of 1L, obtain) repeatedly wash the surface, and the placenta is disinfected;

[0090] (2) Slowly tear the outer layer of amniotic membrane with surgical forceps and place it in a 150mm glass dish, use basic balanced salt solution to repeatedly clean the surface dirty blood, and cut it into small pieces of amniotic membrane tissue with a diameter of 5mm; use a 300-mesh filter Filter, flush the residual blood with normal saline, and transfer the tissue block to a 50ml centrifuge tube;

[0091] (3) ...

Embodiment 2

[0096] Example 2: Preparation of human amniotic mesenchymal stem cells from placental amniotic membrane

[0097] (1) Take out the placenta from the placenta sample collection box, place it in a white porcelain dish, and use a basic balanced salt solution (0.4g of KCl, 0.06g of KH 2 PO 4 , 0.132g of Na 2 HPO 4 .12H 2 O, 8g of NaCl, 0.35g of NaHCO 3 , the D-glucose of 1.0g, the streptomycin of 0.10g, the penicillin of 0.06g are dissolved in water and be settled into the solution of 1L, obtain) repeatedly wash the surface, and the placenta is disinfected;

[0098] (2) Slowly tear the outer layer of amniotic membrane with surgical forceps and place it in a 150mm glass dish, use basic balanced salt solution to repeatedly clean the surface dirty blood, and cut it into small pieces of amniotic membrane tissue with a diameter of 5mm; use a 300-mesh filter Filter, flush the residual blood with normal saline, and transfer the tissue block to a 50ml centrifuge tube;

[0099](3) T...

Embodiment 3

[0104] Example 3: Preparation of human amniotic mesenchymal stem cells from placental amniotic membrane

[0105] (1) Take out the placenta from the placenta sample collection box, place it in a white porcelain dish, and use a basic balanced salt solution (0.4g of KCl, 0.06g of KH 2 PO 4 , 0.132g of Na 2 HPO 4 .12H 2 O, 8g of NaCl, 0.35g of NaHCO 3 , the D-glucose of 1.0g, the streptomycin of 0.10g, the penicillin of 0.06g are dissolved in water and be settled into the solution of 1L, obtain) repeatedly wash the surface, and the placenta is disinfected;

[0106] (2) Slowly tear the outer layer of amniotic membrane with surgical forceps and place it in a 150mm glass dish, use basic balanced salt solution to repeatedly clean the surface dirty blood, and cut it into small pieces of amniotic membrane tissue with a diameter of 5mm; use a 300-mesh filter Filter, flush the residual blood with normal saline, and transfer the tissue block to a 50ml centrifuge tube;

[0107] (3) ...

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Abstract

The invention relates to a method for preparing amniotic mesenchymal stem cells from a human placenta amnion and application of the amniotic mesenchymal stem cells. The method comprises the following steps: washing and disinfecting a placenta; washing and disinfecting an amnion and crushing the amnion; carrying out tissue digestion; carrying out cell culture on a filtrate part and a tissue block part; harvesting the cells; counting and determining the survival rate of the cells, and cryopreserving. The invention further relates to the amniotic mesenchymal stem cells prepared by the method and therapeutic application of the amniotic mesenchymal stem cells. The method provided by the invention has the advantages showed in the description.

Description

technical field [0001] The invention belongs to the technical field of stem cell therapy, and relates to a method for preparing amniotic mesenchymal stem cells from human placenta amniotic membrane, and also relates to amniotic mesenchymal stem cells prepared by the method, and their therapeutic use. Background technique [0002] Stem cells are the basis for the development of regenerative medicine, and their effective application is an effective way to realize the ultimate dreams of human health and longevity, anti-cancer and anti-aging, eternal youth, and continuous improvement of quality of life. In recent years, embryonic stem cells and adult stem cells have received much attention in the fields of cell biology and regenerative medicine research. However, despite the totipotency of embryonic stem cells that other cells cannot match, factors such as tumorigenicity, immune rejection after allogeneic transplantation, and unavoidable ethical disputes largely limit its clinic...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0775A61K35/28A61P9/10A61P3/10A61P25/00
Inventor 许晓椿王正肖海蓉
Owner BOYALIFE
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