Composition, inducer containing composition and induction method

A technology of composition and preparation, applied in the field of induction preparation and induction, can solve problems such as donor injury

Inactive Publication Date: 2017-04-19
GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, the neuron-like cells induced to differentiate can not effectively and stably express the characteristics of nerve cells. Whether such neuron-like cells have t

Method used

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  • Composition, inducer containing composition and induction method
  • Composition, inducer containing composition and induction method
  • Composition, inducer containing composition and induction method

Examples

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Effect test

Embodiment 1

[0055] Example 1 Primary isolation and culture of GMSCs

[0056] 4) Source of raw materials: The gingiva cut out by clinical orthodontic eruption or impacted tooth extraction is used to remove the gingival tissue surrounding the permanent teeth. Patients are required to have no gingival hyperplasia, inflammation and use of drugs that cause gingival hyperplasia. The excised gingival tissue was quickly immersed in 4°C pre-cooled PBS containing 3 times double antibody.

[0057] Among them, the 3-fold double antibody concentration was 300u / mL penicillin and 300u / mL streptomycin.

[0058] 5) Rinse: Rinse 3 times with PBS containing 3 times the double antibody, and then soak the gingiva in the serum-free medium of mesenchymal stem cells for 5 minutes;

[0059] Among them, the 3-fold double antibody concentration was 300u / mL penicillin and 300u / mL streptomycin.

[0060] Wherein, the serum-free medium of mesenchymal stem cells contains 100u / mL penicillin and 100u / mL streptomycin. ...

Embodiment 2

[0081] Embodiment 2 GMSCs differentiation

[0082] In order to verify the effect of differentiation induction of the present invention, set 4 groups of control groups and 3 groups of experimental groups at the same time, select the 3rd generation GMSCs, press 1×10 4 The cells / mL density was inoculated in a six-well plate with polylysine-treated sterile coverslips, and 2 mL of DMEM / F12 medium containing 10% FBS was added to each well. After 24 hours, replace the culture medium corresponding to each group in the control group 1 ~ control group 4 and the experimental group 1 ~ experimental group 3, at 37 ° C, 5% CO 2 , Culture in saturated humidity, and replace with new culture medium every 2 to 3 days.

[0083] Control group 1 is a negative control group, which is a conventional culture group, and the culture medium used is: DMEM / F12 culture medium containing 10% FBS by volume.

[0084] Control group 2 is a positive control group, and the induction medium used is: DMEM / F12 cultu...

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Abstract

The invention relates to the field of cells, especially to a composition, an inducer containing the composition and an induction method. The composition comprises B27, a basic fibroblast growth factor, an epidermal growth factor and a nerve growth factor. According to the induction method, the B27, the basic fibroblast growth factor (bFGF), the epidermal growth factor (EGF) and the nerve growth factor (NGF) are mainly utilized to induce gingiva mesenchymal stem cell differentiation into neuron-like cells which have a typical morphology of nerve cells and express neuronal marker nestin, neuronspecific enolase and neurogliocyte marker glial fibrillary acidic protein. It shows that the gingiva mesenchymal stem cells retain the capability of transdifferentiation of germinal layers into non-mesenchymal cells, can transdifferentiate germinal layers into neuron-like cells and are expected to become seed cells for nerve cell replacement therapy.

Description

technical field [0001] The invention relates to the field of cells, in particular to a composition, an induction preparation containing the composition and an induction method. Background technique [0002] The recovery of nerve function after central nervous system injury is very difficult, mainly due to the extremely poor regeneration ability of neurons in the brain. Current studies have proved that although neural stem cells exist in the brain and can differentiate into neurons and glial cells, it is still very difficult to use them for repairing neuron damage due to the limited location and very small number of neural stem cells. of. With the deepening of research in the field of stem cells, it has been found that stem cells in other parts can also differentiate into nerve cells. Transplantation of embryonic nerve cells or umbilical cord blood stem cells into the injured mouse brain can moderately improve its neurological function, but due to ethics, immune rejection a...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12N5/0775
CPCC12N5/0619C12N2501/11C12N2501/115C12N2501/13C12N2506/1392
Inventor 葛啸虎陈海佳王一飞戚康艺
Owner GUANGZHOU SALIAI STEMCELL SCI & TECH CO LTD
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