Isolated culture method of human amnia mesenchymal stem cells

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...

AI Technical Summary

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 critical role in the separation and activity of cells. The existing technology uses trypsin and collagenase to separate the separation method. It has the disadvantages of long time-consuming, cumbersome steps, and low cell viability; and the addition of animal-derived components such as fetal bovine serum to the culture medium increases the cost of culture, and also introduces exogenous animal protein, which makes the cultured cells unfavorable for clinical application

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