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Method for inducing neural stem cells to differentiate into neurons and astrocytes by umbilical cord mesenchymal stem cells

A technology of glial cells and neural stem cells, applied in the biological field, can solve the problems of low induction rate of neural stem cells, low success rate, and lack of differentiation function of neural stem cells, and achieve the effect of improving the induction rate

Active Publication Date: 2021-05-14
TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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Problems solved by technology

[0004] Studies have shown that umbilical cord can be used as an ideal source of mesenchymal stem cells to induce neural stem cells from umbilical cord, but the induction rate of umbilical cord mesenchymal stem cells into neural stem cells is low , and the success rate is also low, and the induced neural stem cells do not have differentiation function

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  • Method for inducing neural stem cells to differentiate into neurons and astrocytes by umbilical cord mesenchymal stem cells
  • Method for inducing neural stem cells to differentiate into neurons and astrocytes by umbilical cord mesenchymal stem cells
  • Method for inducing neural stem cells to differentiate into neurons and astrocytes by umbilical cord mesenchymal stem cells

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[0055] The present invention will be further described in detail below through the specific examples, the following examples are only descriptive, not restrictive, and cannot limit the protection scope of the present invention with this.

[0056] A method for inducing neural stem cells to differentiate into neurons and astrocytes by umbilical cord mesenchymal stem cells, the specific method steps are as follows:

[0057] (1) Take the third-generation human umbilical cord mesenchymal stem cells and inoculate them on the culture flask at a density of 3000. The third-generation human umbilical cord mesenchymal stem cells are used from the primary culture to the third generation or commercially available for resuscitation;

[0058] The resuscitation method is as follows: take human umbilical cord mesenchymal stem cells out of the liquid nitrogen tank, resuscitate them with sterile water at 37°C, put them into fresh DF12 medium, the ratio of medium to cell suspension is 9:1, 300G, 5...

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Abstract

The invention relates to a method for inducing neural stem cells to be differentiated into neurons and astrocytes by umbilical cord mesenchymal stem cells. The method comprises the following steps: (1) culturing the third generation of human umbilical cord mesenchymal stem cells; (2) co-culturing double antibodies by using DF12 and 2% serum for 24 hours, and then carrying out flow cytometry; (3) co-culturing and inducing into neural stem cells by using 2% of serum, 2% of B27, DF12, 20 ng / mL of EGF, 20 ng / mL of BFGF, 10 [mu] M of Forskolin, 1 mM of IBMX and 35 ng / mL of dbcAMP, and then adding differentiation factors; (4) detecting expressions of markers of four neural stem cells, namely Netin, PAX6, SOX1 and SOX2, by using immunofluorescence and fluorescent quantitative PCR; and (5) detecting the expression of the glial cell marker GFAP and the neuron marker MAP-2 by using an immunofluorescence technology and a fluorescent quantitative PCR technology. According to the invention, multiple factors are used in a combined manner, the addition amount is optimized, neurons and astrokeratinocytes can be differentiated at the same time, the marker is highly expressed, and the induction rate of umbilical cord mesenchymal stem cells to induce neural stem cells is improved, so that the neural stem cells can be obtained to be used for treating neuronal injury diseases.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a technique for inducing umbilical cord mesenchymal stem cells, in particular to a method for inducing neural stem cells to differentiate into neurons and astrocytes by umbilical cord mesenchymal stem cells. Background technique [0002] As the people's life in our country is getting richer and the aging population is becoming more prominent, more and more elderly people are suffering from nerve damage diseases, such as Parkinson's disease, Alzheimer's disease, and cerebral thrombosis and other local nerve damage and apoptosis diseases. At present, there is still no effective clinical treatment method to improve the neurological deficit of patients, and it can only relieve the symptoms or delay the exacerbation of the disease. Experiments have proved that neural stem cell transplantation can improve the recovery of neuron damage diseases. [0003] At present, neural stem cells...

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Application Information

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IPC IPC(8): C12N5/079C12N5/0793
CPCC12N5/0622C12N5/0619C12N2502/1388
Inventor 冯郸马洁楼敏铭崔瑛
Owner TIANJIN KANGTING BIOLOGICAL ENG GRP CO LTD
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