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Culture medium for inducing mesenchymal stem cells to be differentiated into nerve cells

A technology of nerve cells and stem cells, applied in the culture process, tissue culture, animal cells, etc., can solve the problems of low cell viability and high death rate of cell aberration, and achieve the effect of reducing the number of cell aberration and death

Inactive Publication Date: 2017-03-15
浙江译美生物科技有限公司
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Problems solved by technology

[0003] For example, the patent with the notification number CN101117625 provides a method for inducing differentiation of mesenchymal stem cells and adipose stem cells into neurons and a special medium for inducing the differentiation of mesenchymal stem cells or adipose stem cells into neurons, It is a DMEM medium supplemented with 8-12% fetal bovine serum by volume / volume percentage, but during the actual induction process of this medium, the cell distortion and death rate are high, resulting in low cell viability

Method used

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  • Culture medium for inducing mesenchymal stem cells to be differentiated into nerve cells
  • Culture medium for inducing mesenchymal stem cells to be differentiated into nerve cells
  • Culture medium for inducing mesenchymal stem cells to be differentiated into nerve cells

Examples

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Embodiment 1-6

[0024] Embodiment 1-6: A medium for inducing mesenchymal stem cells to differentiate into nerve cells, which is prepared by the following method:

[0025] Add tretinoin, 2-mercaptoethanol, folic acid, gentamicin, and fluoroquinolone to the basal medium, mix evenly, pass through the membrane to remove bacteria, and obtain a mesenchymal stem cell medium in Examples 1-6. Each component is shown in Table 1.

[0026] The composition and content thereof of table 1 embodiment 1-6

[0027]

Embodiment 7

[0029] Dental pulp mesenchymal stem cells, bone marrow mesenchymal stem cells, umbilical cord mesenchymal stem cells and adipose mesenchymal stem cells were divided into 5000 cells / cm 2 The density was planted in the T75 cell culture flask filled with the medium 1-6 in the above-mentioned examples at 37°C, 6% CO 2 Cultured in a sterile incubator, the medium was changed every 2 days, harvested after 14 days of culture, neuron-specific markers (NSE and NF) and astrocyte-specific markers (GFAP) in the culture medium ) to perform immunocytostaining chemical determination and use the Invitrogen company's automatic cell counter Countess to measure the cell number and cell viability. The test results are shown in Table 2 and Table 3.

[0030] Table 2: Cell staining analysis results of neuron-specific markers

[0031]

[0032] Table 3: Detection results of cell number and cell viability

[0033]

[0034] It can be seen from Table 2 that when the medium contains at least two of...

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Abstract

The invention discloses a culture medium for inducing mesenchymal stem cells to be differentiated into nerve cells. The culture medium comprises a basal culture medium and an inductive agent added in the basal culture medium, wherein the inductive agent is prepared from vitamin A acid, 2-mercaptoethanol and folic acid; in addition, in the culture medium for inducing the mesenchymal stem cells to be differentiated into the nerve cells, the concentration of the vitamin A acid is 10 to 50mu g / ml, the concentration of the 2-mercaptoethanol is 60 to 95mu g / ml, and the concentration of the folic acid is 65 to 75mu g / ml. According to the culture medium, the distortion and death number of cells in an inducing process can be effectively reduced, and higher surviving rate of the nerve cells formed by inducing is guaranteed.

Description

technical field [0001] The invention relates to the field of stem cell differentiation culture, in particular to a culture medium for inducing mesenchymal stem cells to differentiate into nerve cells. Background technique [0002] Neuron damage diseases (such as Parkinson's disease, Alzheimer's disease, cerebral ischemia, etc.) are major diseases that currently plague human beings. At present, there is no drug that can cure neuron damage diseases. Some drugs used to treat the disease Symptoms can only be alleviated to some extent, and long-term use can also produce toxic side effects. In recent years, the use of stem cell transplantation to treat neuron damage diseases has received extensive attention from the medical community. Mesenchymal stem cells (MSCs) are highly expected due to their easy acquisition, low immunogenicity, strong proliferation ability, multidirectional differentiation potential, and few ethical controversies. [0003] For example, the patent with the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/079
CPCC12N5/0618C12N2500/30C12N2500/38C12N2500/44C12N2506/1346
Inventor 汪铮吴振化
Owner 浙江译美生物科技有限公司
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