Method for inducing differentiation of neural crest stem cells towards neurons

A neuron cell and stem cell technology, applied in nervous system cells, biochemical equipment and methods, animal cells, etc., can solve the problems of complex operation and low proportion of neuronal cell differentiation, and achieve simple and convenient operation, risk avoidance and immunity. Originality, the effect of increasing the differentiation ratio

Inactive Publication Date: 2017-07-04
周婧
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AI Technical Summary

Problems solved by technology

These methods are more complicated to operate, and the proportion of neuron cell differentiation is not high

Method used

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Experimental program
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Effect test

Embodiment 1

[0009] Clean the surface of the strontium titanate {110} crystal surface single wafer with deionized water in ultrasonic wave; irradiate the cleaned strontium titanate {110} crystal surface single wafer under ultraviolet light (wavelength 254nm, power 16W), The irradiation time was 20min; the ratio of terminal hydroxyl group / bridging hydroxyl group formed in situ on the surface of strontium titanate {110} crystal plane was 2.0; the neural crest stem cells were divided into 5×10 4 / cm 2 The density of strontium titanate was evenly planted on a single chip of strontium titanate {110} crystal surface, cultured in conventional DMEM basal medium, and the medium was changed every two days; after 14 days of culture, mature and differentiated cells were obtained by flow cytometry identification. The proportion of neuronal cells was 67%.

Embodiment 2

[0011] Clean the surface of the strontium titanate {110} crystal surface single wafer with deionized water in ultrasonic wave; irradiate the cleaned strontium titanate {110} crystal surface single wafer under ultraviolet light (wavelength 254nm, power 16W), The irradiation time was 30min; the ratio of terminal hydroxyl group / bridging hydroxyl group formed in situ on the surface of strontium titanate {110} crystal plane was 2.3; the neural crest stem cells were divided into 5×10 4 / cm 2 The density of strontium titanate was evenly planted on a single chip of strontium titanate {110} crystal surface, cultured in conventional DMEM basal medium, and the medium was changed every two days; after 14 days of culture, mature and differentiated cells were obtained by flow cytometry identification. The proportion of neuronal cells was 78%.

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Abstract

The invention discloses a method for inducing differentiation of neural crest stem cells towards neurons. Firstly, single-crystal surface of a strontium titanate {110} crystal plane is subjected to radiation treatment by virtue of ultraviolet light, so as to special a surface having special hydroxyl properties; and then, the neural crest stem cells are cultivated on the surface by virtue of a conventional DMEM basal culture medium for 14 days, so as to obtain the neurons which are mature in differentiation. The method, which induces the directional differentiation of the stem cells on the basis of the hydroxyl properties of the material surface, provided by the invention is simple and convenient to operate and is quite low in cost, and a differentiation proportion of the neural crest stem cells towards the neurons can be obviously improved; and moreover, risks and immunogenicity caused by application of animal serum and biochemical reagents can be avoided; therefore, the invention is applicable to such fields as treatment of neurodegenerative disease and the like.

Description

technical field [0001] The invention relates to a method for inducing neural crest stem cells to differentiate into neuron cells. Background technique [0002] Stem cell directional differentiation regulation is one of the key topics in stem cell research. In addition to biochemical methods such as gene transcription factors, cell growth factors, and chemical inducers, it has been found that the surface properties of materials play a decisive role in mediating the directed differentiation of stem cells in vitro. A new hotspot of research. Neural crest stem cells can differentiate into neurons, astrocytes, oligodendrocytes, pigment cells, and odontocytes. Mediating the directed differentiation of neural crest stem cells into neurons is an important basis for the use of stem cell therapy to treat neurodegenerative diseases. Patent applications CN201380080663.2, CN201380080656.2, and CN201010163969.4 all use the addition of biochemical reagents or biological factors for dire...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0793
CPCC12N5/0619C12N2506/08C12N2533/20
Inventor 周婧
Owner 周婧
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