Method for differentiation of amniotic fluid mesenchymal stem cells into neural stem cells

Abstract

The invention provides a method for differentiation of amniotic fluid mesenchymal stem cells into neural stem cells, and the method comprises the following steps: centrifugally separating stem cells in amniotic fluid tissues to obtain the amniotic fluid mesenchymal stem cells; culturing the amniotic fluid mesenchymal stem cells by using an amniotic fluid mesenchymal stem cell serum-free proliferation culture medium; performing passage to P3-generation amniotic fluid mesenchymal stem cells; digesting the P3-generation amniotic fluid mesenchymal stem cells by using trypsin to obtain a single-cell suspension, and culturing the single-cell suspension overnight; removing the amniotic fluid mesenchymal stem cell serum-free proliferation culture medium, and then cleaning, and then adding the cells into a first induction culture medium for induction culture; and removing the first induction culture medium, then digesting the cells with trypsin to obtain a single-cell suspension, and carrying out inoculation culture on the single-cell suspension by using a second induction culture medium to obtain the cell ball formed by the neural stem cells. According to the invention, a variety of specific cell factors are used, and the amniotic fluid mesenchymal stem cells are induced by a two-step method to obtain the neural stem cells, wherein the obtained neural stem cells have the ball formationgrowth characteristics of neural stem cells.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, in particular to a method for differentiating amniotic fluid mesenchymal stem cells into neural stem cells. Background technique [0002] Stem cells are a type of primitive undifferentiated cells with self-replication and multi-directional differentiation capabilities, have strong proliferative activity, and can differentiate into mature cells of different germ layers under specific conditions, such as epidermal cells, nervous system cells, liver cells, bone cells Wait. [0003] The incidence of central nervous system damage or degenerative diseases such as Parkinson's (PD) and stroke is increasing day by day, becoming an important factor that threatens human health. Neural stem cells differentiate into a variety of functional neurons and become important seed cells for the treatment of nervous system damage and degenerative diseases. At present, neural stem cells are mainly i...

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Problems solved by technology

[0007] However, most of the nerve-like cells commonly used at this stage are induced by adding chemical inducers such as β-mercaptoethanol, DMSO, etc. Although they have the cell morphology of nerve-like cells after induction, some studies have suggested that this is only because of the cytotoxicity of chemical reagents. resulting in morphological changes, the resulting nerve-like cells do not have the unique functions of nerve cells

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