A method for preparing chorionic mesenchymal stem cells

Abstract

The invention provides a method for preparing chorionic mesenchymal stem cells, comprising aseptically collecting placenta; isolating chorionic tissue; obtaining chorionic mesenchymal stem cells; cultivating chorionic mesenchymal stem cells; Cryopreservation; recovery of chorionic mesenchymal stem cells. The method of the present invention has the following technical effects: the pollution prevention method reduces the probability of pollution from the source of collection, and the surface is washed multiple times to effectively reduce the probability of pollution; the pollution of mixed cells is reduced; the process can be simplified by using a single enzyme for digestion; the use of serum-free medium for cultivation Reduce the use of animal-derived components, and the cell performance is stable, which can maintain the long-term culture process of chorionic mesenchymal stem cells in vitro, maintain cell shape, proliferation ability, expression of MSC surface markers, differentiation ability, and maintain chorionic mesenchymal stem cells in vitro During the long-term culture process, the expression of telomerase, the stable expression of oncogenes, and the stable karyotype are maintained; enzyme digestion and cryopreservation recovery do not harm the cells.

Description

technical field [0001] The invention relates to a method for preparing chorionic 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, compact bone, cartilage, muscle, umbilical cord, and placenta. Mesenchymal cells isolated from placental tissue can differentiate into a variety of cells with specific functions under different conditions, such as osteoblasts, chondrocytes, adipocytes, cardio...

AI Technical Summary

Problems solved by technology

[0005] 1) The existing technologies of placenta collection and chorion separation lack effective pollution reduction measures, and the anti-pollution measures have limited effect. The pollution rate of primary culture is high, and there is no method to reduce pollution from the source. Inhibiting the reproduction of contaminating bacteria during the process cannot effectively reduce pollution; adding antibiotics to the culture system is not conducive to the clinical application of chorionic stem cells

[0006] 2) The existing enzymatic digestion method is complex and requires multiple enzymatic digestions;

[0007] 3) Operations such as enzyme digestion, cryopreservation and recovery may cause cell damage to the cells;

[0008] 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 chorionic mesenchymal stem cells, prevent cell aging, and maintain cell proliferation Retain differentiation capacity during the process;

[0009] 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 chorionic mesenchymal stem cells

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