Method for preparing stem cells from human amniotic membrane

Abstract

The invention discloses a method for preparing stem cells from a human amniotic membrane, specifically relates to a method for separating epithelial cells and mesenchymal stem cells from the amniotic membrane, and relates to the technical field of cell separation. The method comprises the following steps: collecting a placenta by self-made sterile, efficient and stable placenta protecting liquid; and separating amniotic membrane epithelial cells and amniotic membrane mesenchymal stem cells by combining a digestion process and a climbing process to respectively obtain high-purity amniotic membrane epithelial cells and high-purity amniotic membrane mesenchymal stem cells. The method disclosed by the invention solves a problem that subsequent cell separation and culture is directly affected by high human amniotic membrane collecting pollution rate and poor activity of the collected cells in the prior art. According to the method disclosed by the invention, the method for separating the stem cells from the amniotic membrane is improved and created to form a standard separating scheme, so that the stem cell resource can be utilized more stably and sufficiently, pollution is avoided, purity is high and the number of the cells is large; while the cell activity is kept, original characteristics of the stem cells are ensured.

Description

technical field

[0001] The invention relates to a cell separation technology, in particular to a method for separating epithelial cells and mesenchymal stem cells from amnion. Background technique

[0002] Amnion is the product of embryonic development. Amnion cells have stem cell characteristics and do not express human leukocyte antigens HLA-I and HLA- , There is no allogeneic immune rejection, so it is suitable for cell therapy. The stem cells in the amniotic membrane mainly include epithelial cells and mesenchymal stem cells.

[0003] Amnion epithelial cells have the characteristics of subtotipotent stem cells, have strong differentiation ability, can differentiate into three germ layers, and have no tumorigenicity and ethical problems. They are ideal seed cells for tissue engineering and cell therapy. Amniotic mesenchymal stem cells have a certain multi-directional differentiation potential, but compared with amniotic epithelial cells, their main feature is that they ...

Examples

Embodiment Construction

[0121] The present invention will be further described below in conjunction with accompanying drawing and embodiment:

[0122] A method for preparing stem cells from human amniotic membrane, which includes collecting, separating, culturing, freezing, and detecting; wherein, during the separation process, the amniotic membrane epithelial cells are digested with enzymes, and the amniotic membrane mesenchymal stem cells are then sliced ​​by tissue method, the two are completed step by step.

[0123] The collection of the present invention specifically includes: putting the collected placenta into a sterile bag containing D-Hanks liquid placenta protection solution of 0.2 g / L streptomycin and 0.12 g / L penicillin, sealing the sterile bag and putting Put the collection box into the collection box, put the collection box into the constant temperature collection box, and send it to the stem cells within 24 hours with the constant temperature collection box.

[0124] The placenta coll...

technical field

[0001] The invention relates to a cell separation technology, in particular to a method for separating epithelial cells and mesenchymal stem cells from amnion. Background technique

[0002] Amnion is the product of embryonic development. Amnion cells have stem cell characteristics and do not express human leukocyte antigens HLA-I and HLA- , There is no allogeneic immune rejection, so it is suitable for cell therapy. The stem cells in the amniotic membrane mainly include epithelial cells and mesenchymal stem cells.

[0003] Amnion epithelial cells have the characteristics of subtotipotent stem cells, have strong differentiation ability, can differentiate into three germ layers, and have no tumorigenicity and ethical problems. They are ideal seed cells for tissue engineering and cell therapy. Amniotic mesenchymal stem cells have a certain multi-directional differentiation potential, but compared with amniotic epithelial cells, their main feature is that they ...

Examples

Embodiment Construction

[0121] The present invention will be further described below in conjunction with accompanying drawing and embodiment:

[0122] A method for preparing stem cells from human amniotic membrane, which includes collecting, separating, culturing, freezing, and detecting; wherein, during the separation process, the amniotic membrane epithelial cells are digested with enzymes, and the amniotic membrane mesenchymal stem cells are then sliced ​​by tissue method, the two are completed step by step.

[0123] The collection of the present invention specifically includes: putting the collected placenta into a sterile bag containing D-Hanks liquid placenta protection solution of 0.2 g / L streptomycin and 0.12 g / L penicillin, sealing the sterile bag and putting Put the collection box into the collection box, put the collection box into the constant temperature collection box, and send it to the stem cells within 24 hours with the constant temperature collection box.

[0124] The placenta coll...

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