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Application of naringin in promoting proliferation and differentiation of neural stem cell

A technology of neural stem cells, naringin, applied in the fields of regenerative medicine and pharmacology

Active Publication Date: 2014-07-09
北京清美联创干细胞科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] So far, there are no domestic and foreign research reports on the promotion of naringin in promoting the proliferation and differentiation of neural stem cells in vitro

Method used

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  • Application of naringin in promoting proliferation and differentiation of neural stem cell
  • Application of naringin in promoting proliferation and differentiation of neural stem cell
  • Application of naringin in promoting proliferation and differentiation of neural stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Isolation, cultivation and identification of neural stem cells in vitro

[0028] (1) Isolation and culture of neural stem cells derived from midbrain

[0029] E14d mice were killed by necking, the embryos were taken out by aseptic operation, the midbrain tissue of the embryos was separated, digested into single cells, counted with trypan blue, and counted with 2x10 5 cells / mL inoculated in a T25 culture flask at 37°C, 5% CO 2 For suspension culture, the stem cell culture medium is DMEM / F12 (1:1), 2% B27, bFGF (10 ng / mL), EGF (20 ng / mL). Change the medium in half every 3 to 4 days, and observe the number and size of the growing suspended cell spheres after one week of culture. The neurospheres were collected, mechanically blown into a single-cell suspension, and placed under the same conditions to continue culturing, depending on the growth status to determine whether to pass.

[0030] (2) Continuous passage and induced differentiation of cell subclones

[...

Embodiment 2

[0035] Example 2 Proliferation-promoting effect of naringin on neural stem cells

[0036] (1) Experimental design and drug treatment

[0037] The experiment was divided into five groups, namely the control group, 5x10 -7 mol / L naringin group, 5x10 -6 mol / L naringin group, 5x10 -5 mol / L naringin group, 5x10-4 mol / L naringin group. Collect the second-generation neural stem cells from the primary culture, and use 2x10 5 Cells / mL were inoculated in 24-well plates, 500uL per well, and 6 replicate wells were set up for each treatment group. The number of neurospheres and the survival rate of cells were observed after 4 days and 7 days of culture respectively.

[0038] (2) Detection method of neural stem cell proliferation

[0039] ① Neurosphere counting method: After the second-generation neural stem cells cultured in the primary culture were cultured for 4 days and 7 days under the action of different concentrations of naringin, the number of neurospheres formed was counted in...

Embodiment 3

[0043] Example 3 The ability of naringin to induce neural stem cells to differentiate into functional neurons

[0044] (1) Induction and differentiation of neural stem cells by naringin

[0045] The second-generation neural stem cell spheres cultured at the primary stage were mixed with 5x10 -4 mol / L of naringin 7 days after the neural stem cell spheres were collected, blown into a single cell suspension in the differentiation medium (DMEM / F12+10% fetal bovine serum) respectively, with 2x10 5 The concentration of each / ml was inoculated in a 24-well plate on poly-L-lysine-treated glass slides, 500 uL per well, and six replicate wells were set up for each group of treatments. After 24 hours, it can be seen that the cells are all attached to the wall, and different concentrations of naringin (5x10 -6 , 5x10 -5 , 5x10 -4 mol / L) after treatment, the culture plate was placed in saturated humidity, 5% CO 2 , 37 ° C incubator to continue culturing for 7 days.

[0046] (2) Immuno...

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Abstract

The invention belongs to the field of regenerative medicine and pharmacology, relates to an application of a double hydrogen flavonoid small molecule compound, and particularly relates to a new application of naringin in promoting proliferation and differentiation of a neural stem cell. The invention is characterized by relating to an application of the naringin in medicament for neural stem cell transplantation in the treatment of neurodegenerative diseases, an application of the naringin in repairing damaged nerve tissues and preparation a function-reconstructable nerve nutriment and cell differentiation agent, and an application of the naringin in constructing an efficient medicament screening model and applying the efficient medicament screening model to preliminary screening and evaluation of the medicament efficacy. The invention confirms the new pharmacological activity of the naringin, establishes an application foundation in stem cell transplantation (SCT), and provides new thoughts for developing new medicaments with independent intellectual property rights.

Description

technical field [0001] The invention belongs to the field of regenerative medicine and pharmacology, and relates to the use of a dihydroflavonoid small molecule compound, in particular to the new use of naringin in promoting the proliferation and inducing differentiation of neural stem cells in vitro. Background technique [0002] Naringin, also known as naringin, citrusin, and isohesperidin, is mainly found in the peel and pulp of grapefruit, tangerine, and orange of Rutaceae plants. Naringin is also the main active ingredient of Chinese herbal medicine Rhizoma Drynariae, Aurantium Fructus Aurantium, Fructus Aurantium, and Tangerine. The content of naringin in various plants varies greatly with the variety and place of origin, and the content of immature fruit is usually higher. [0003] The chemical structural formula of naringin is as follows: [0004] [0005] Molecular formula: C 27 h 32 o 14 [0006] Molecular weight: 580.54 [0007] Preparation of naringin: ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0797C12N5/0793A61K31/7048A61K35/30A61P25/28C12Q1/02
Inventor 田杰
Owner 北京清美联创干细胞科技有限公司
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