Isolated culture method of human amnia mesenchymal stem cells

A technique for isolating and culturing stem cells, applied in the field of biomedicine, can solve the problems of long culture period, low cell viability, low cell purity, etc., and achieve the effects of uniform and stable phenotype, strong proliferation ability, and short culture period.

Inactive Publication Date: 2015-11-18
GENESIS STEMCELL REGENERATIVE MEDICINE ENG CO LTD
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Problems solved by technology

At present, hAMSCs are separated by two methods: tissue block culture method and enzyme digestion method. The tissue block culture method has the disadvantages of long culture period and low cell purity, which is difficult to meet the needs of stem cell clinical research. Enzyme digestion method is the basis of tissue block culture. It has the advantages of short cell culture period and high cell purity, but the selection of the type of digestive enzyme plays a very c

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  • Isolated culture method of human amnia mesenchymal stem cells
  • Isolated culture method of human amnia mesenchymal stem cells
  • Isolated culture method of human amnia mesenchymal stem cells

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Embodiment 1

[0037] see Figure 1-8 , in an embodiment of the present invention, a method for isolating and culturing human amniotic mesenchymal stem cells, comprising the following steps:

[0038] 1. Preparation of mesenchymal stem cells

[0039] (1) Collection of human placenta and amniotic membrane tissue: After the human placenta is removed, it is immersed in the placenta preservation solution and kept at 2-8°C for no more than 24 hours.

[0040] (2) Disinfection of amniotic membrane tissue: Take the placenta out of the placental preservation solution, place it flat on a sterile tray with the side containing the amniotic membrane tissue facing up, wipe it clockwise with 75% alcohol cotton, repeat twice, and start from the placenta with the umbilical cord as the The sterilized amniotic tissue was stripped centrally.

[0041](3) Washing: Put the amnion tissue into a 90cm sterile petri dish, wash it twice with tissue sample washing solution, with an interval of 5 minutes, and then wash ...

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Abstract

The invention discloses an isolated culture method of human amnia mesenchymal stem cells, which comprises the following steps: carrying out human placenta amnia tissue acquisition, amnia tissue disinfection, washing, shredding and mixed collagenase digestion to obtain mesenchymal stem cells, and carrying out mesenchymal stem cell in-vitro culture. The method can obtain abundant mesenchymal stem cells, which have the advantages of uniform and stable phenotype and high multiplication capacity and have the multidirectional differentiation potential, from the human amnia tissues; and the culture medium in-vitro amplification is performed to purify the cells, thereby satisfying the quantity for cryopreservation or clinical use. The method has the advantages of short cell culture period and high repetitiveness. The stem cells obtained by the method have the phenotypes of CD105<+>, CD90<+> and CD73<+>.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a method for separating and culturing human amniotic mesenchymal stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of primitive cells with self-renewal ability and multilineage differentiation potential under suitable microenvironment. It is first found in bone marrow and subsequently isolated in fat, bone, muscle, lung, liver, pancreas, and in umbilical cord, cord blood, and placental tissue. Because of its strong proliferation ability, strong differentiation ability, low immunogenicity and immune regulation function, its clinical application value has attracted more and more attention. [0003] As a cell product to be used in clinical cell therapy, it should have the characteristics of convenient tissue collection, wide sources, and no ethical restrictions; simple isolation process, etc., while bone marrow-derived MSCs are difficult to obtain, and th...

Claims

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Application Information

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IPC IPC(8): C12N5/0735C12N5/0775
Inventor 王军霞殷鉴强
Owner GENESIS STEMCELL REGENERATIVE MEDICINE ENG CO LTD
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