A method for preparing decidual mesenchymal stem cells

Abstract

The invention provides a method for preparing decidual mesenchymal stem cells, comprising using male full-term fetal placenta as raw material, aseptically treating the placenta; isolating decidual tissue; identifying decidual tissue and obtaining decidual mesenchymal stem cells from Identification of maternal tissue; culture of decidual mesenchymal stem cells; cryopreservation of decidual mesenchymal stem cells; recovery of decidual mesenchymal stem cells. The method of the present invention has the following technical features: collect the placenta where the fetus is male, and determine whether it comes from the mother through sex identification after obtaining the decidua tissue without contamination from the daughter tissue; prevent bacterial contamination, reduce the probability of contamination from the source of collection, and wash it multiple times The surface can effectively reduce the chance of contamination; the use of a single enzyme for digestion can simplify the process; the use of serum-free medium for culture reduces the use of animal-derived components, and the cell performance is stable, which can maintain the long-term culture of decidual mesenchymal stem cells in vitro, maintain cell shape, Proliferation ability, expression of MSC surface markers, differentiation ability, etc.

Description

technical field [0001] The invention relates to a method for preparing decidual mesenchymal stem cells, belonging to the technical field of stem cells. Background technique [0002] Mesenchymal stem cells (MSCs) are a type of pluripotent stem cells derived from the developing mesoderm, which have the potential of self-renewal and multilineage differentiation. Form bone, cartilage, fat, cardiac muscle and other tissues. Mesenchymal stem cells were originally mainly derived from bone marrow, but the content of mesenchymal stem cells (BMMSC) in human bone marrow is extremely low. In recent years, mesenchymal cells with stem / progenitor cell characteristics have been isolated from various tissues such as peripheral blood, dental pulp, cartilage, muscle, umbilical cord, and placenta. Umbilical cord mesenchymal stem cells, amniotic mesenchymal stem cells and chorion mesenchymal stem cells derived from the daughter body can be isolated from the placenta tissue, as well as decidual...

AI Technical Summary

Problems solved by technology

[0005] 1) The existing technology of decidua tissue separation cannot remove the pure decidual tissue from the mother, and the chorionic tissue or amniotic tissue from the daughter that is always mixed with the placenta

[0006] 2) The existing technologies for placenta collection and decidua separation lack effective pollution reduction measures, and the anti-pollution measures have limited effect. Inhibiting the reproduction of contaminating bacteria during the cultivation process cannot effectively reduce pollution;

[0007] 2) The existing enzymatic digestion method is complex and requires multiple enzymatic digestions;

[0008] 3) Operations such as enzyme digestion, cryopreservation and recovery may cause cell damage to the cells;

[0009] 4) In the process of stem cell culture, cells are prone to aging, flat cells, slow proliferation or stop proliferation, loss of differentiation ability, etc. At present, there is no method or technical experience to effectively maintain the shape of decidual mesenchymal stem cells, prevent cell aging, and maintain cell proliferation Retain differentiation capacity during the process;

[0010] 5) After long-term passage of mesenchymal stem cells, there may be risks such as inactivation of telomerase, increased expression of oncogenes, decreased expression of tumor suppressor genes, and changes in karyotype. Currently, there is a lack of research data on decidual mesenchymal stem cells.

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