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Method for inducing differentiation from neuroblastoma cells to neurons

A technology for neuroblastoma and neurons, applied in the field of cell transdifferentiation, can solve the problems of high cost, long differentiation time, low neuron cell ratio, etc., achieve ratio improvement, synapse growth improvement, increase ratio and success rate effect

Pending Publication Date: 2020-07-28
THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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Problems solved by technology

[0004] Through the above analysis, the existing problems and defects of the prior art are: the existing method for differentiating neuroblastoma cells into neurons, the ratio of neuron cells produced by differentiation of neuroblastoma cells is very low, and the differentiation time is relatively long ,higher cost

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  • Method for inducing differentiation from neuroblastoma cells to neurons
  • Method for inducing differentiation from neuroblastoma cells to neurons
  • Method for inducing differentiation from neuroblastoma cells to neurons

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Experimental program
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Embodiment

[0071] Example: Differentiation Rate

[0072] In order to identify the differentiation rate of neurons formed by inducing the differentiation of neuroblastoma cells, the present invention performs cell immunofluorescent chemical staining on the cultured cells for the neuron-specific marker β-tubulin III.

[0073] On the third and seventh days of induction of differentiation, the coverslips with grown cells were washed twice with PBS and fixed with 4% paraformaldehyde (Sigma) for 30 minutes at room temperature. After washing twice with PBS, cells were blocked with PBS containing 5% goat serum for 1 hour at room temperature, and then incubated overnight at 4°C with rabbit anti-β-tubulin III antibody (1:1000, Abcam). After the cells were washed twice with PBS the next day, they were incubated with TRITC-conjugated goat anti-rabbit IgG antibody (1:200, Jackson ImmunoResearchLab) for 2 hours at room temperature in the dark. Finally, cells were stained with DAPI (VectorLabs) and pl...

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Abstract

The invention belongs to the technical field of cell transdifferentiation, and discloses a method for inducing differentiation from neuroblastoma cells to neurons. The method comprises the following steps of acquiring the neuroblastoma cells, performing incubation and dispersion treatment to obtain single cell suspension liquid, performing centrifugation for removing supernate, precipitating the cells, adding a culture solution, performing uniform mixing, and performing subculture; and performing differentiation in a differentiation culture medium to form neuroblastoma cell-neurospheres, performing crushing, performing inoculation into a culture container containing an induction culture medium, and performing differentiation culture for 3-7 days to obtain neuronal cells. According to the method for inducing differentiation from the neuroblastoma cells to the neurons, differentiation induced formation of the neurons can be accelerated, so that the proportion of the neuronal cells generated by differentiation of the neuroblastoma cells is greatly increased. Experiments show that on the third day of induction, the neuron proportion is higher than 80%, on the seventh day, the neuron proportion reaches 95%, and the cell maturity and the neurosynaptic growth of the neuronal cells are also improved.

Description

technical field [0001] The invention belongs to the technical field of cell transdifferentiation, and in particular relates to a method for inducing neuroblastoma cells to differentiate into neurons. Background technique [0002] Neuroblastoma is currently the most common extracranial tumor in children and the most common tumor in infants and young children. Nearly half of neuroblastomas occur in infants and young children under the age of 2. Neuroblastoma accounts for about 6-10% of childhood tumors, and 15% of childhood tumor mortality. For children under 4 years old, the death rate per million population is 10; for children 4 to 9 years old, the death rate per million population is 4. Neuroblastoma is a neuroendocrine tumor that can arise from any neural crest of the sympathetic nervous system. It most commonly occurs in the adrenal glands, but can also occur in the nervous tissue of the neck, chest, abdomen, and pelvis. Neuroblastoma cell differentiation into neurona...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12M3/00C12M1/36C12M1/38C12M1/33C12M1/12C12M1/00
CPCC12N5/0619C12M41/22C12M41/34C12M41/00C12M33/10C12M33/14C12N2506/30C12N2509/00
Inventor 鹿洪亭刘伟强李富江陈伟明
Owner THE AFFILIATED HOSPITAL OF QINGDAO UNIV
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