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Method for enhancing differentiation of mesenchymal stem cells into nerve cells

A technology of nerve cells and mesenchymal stem cells, applied in biochemical equipment and methods, animal cells, nervous system cells, etc., can solve the limited number of mesenchymal stem cells, the loss of mesenchymal stem cell capacity, and the difficulty of clinical application of mesenchymal stem cells and other problems, to achieve the effect of simple and easy-to-learn operation process and solve the effect of proliferation and directional differentiation into nerve cells

Inactive Publication Date: 2019-01-29
华子昂 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the number of mesenchymal stem cells in the human body is limited, and mesenchymal stem cells have the problem of gradual loss of ability in the process of passage and proliferation in vitro. This problem has caused great difficulties to the clinical application of mesenchymal stem cells, so it is necessary to For the success of stem cell therapy for nervous system diseases, the proliferation and differentiation of seed stem cells must first be solved

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Take the human bone marrow mesenchymal stem cells that have been subcultured to the third passage and grow well, and digest them with 0.25% trypsin to form single cells in suspension, and use 4x10 4 / mL density to spread bone marrow mesenchymal stem cells into 6-well cell culture plates.

[0024] (2) Bone marrow mesenchymal stem cell medium: 10.0% FBS, 2mmol L -1 L-Glutamine, 100U·mL -1 Streptomycin, 100U·mL -1 Penicillin-based αMEM medium, sheep placenta extract was added to each culture well to a final concentration of 0.4 μg / mL, and the culture conditions were: 37°C, 5% CO 2 cultured in a cell culture incubator. Monitor cell growth status by MTT method: remove the medium, wash with PBS buffer 3 times, add 20 μL of MTT reagent to each duplicate well, continue to stand in the cell culture incubator for 4 hours, remove the medium again, wash with PBS After washing with buffer solution for 3 times, 150 μL of DMSO reagent was added to each duplicate well, and the ...

Embodiment 2

[0028] (1) Take the human bone marrow mesenchymal stem cells that have been subcultured to the third passage and grow well, and digest them with 0.25% trypsin to form single cells in suspension, and use 4x10 4 / mL density to spread bone marrow mesenchymal stem cells into 6-well cell culture plates.

[0029] (2) Bone marrow mesenchymal stem cell medium: 10.0% FBS, 2mmol L -1 L-Glutamine, 100U·mL -1 Streptomycin, 100U·mL -1 Penicillin-based αMEM medium, sheep placenta extract was added to each culture well to a final concentration of 0.4 μg / mL, and the culture conditions were: 37°C, 5% CO 2 cultured in a cell culture incubator. Monitor cell growth status by MTT method: remove the medium, wash with PBS buffer 3 times, add 20 μL of MTT reagent to each duplicate well, continue to stand in the cell culture incubator for 4 hours, remove the medium again, wash with PBS After washing with the buffer solution for 3 times, 150 μL of DMSO reagent was added to each duplicate well, and ...

Embodiment 3

[0033] (1) Take the human bone marrow mesenchymal stem cells that have been subcultured to the third passage and grow well, and digest them with 0.25% trypsin to form single cells in suspension, and use 4x10 4 / mL density to spread bone marrow mesenchymal stem cells into 6-well cell culture plates.

[0034] (2) Bone marrow mesenchymal stem cell medium: 10.0% FBS, 2mmol L -1 L-Glutamine, 100U·mL -1 Streptomycin, 100U·mL -1 Penicillin-based αMEM medium, sheep placenta extract was added to each culture well to a final concentration of 0.4 μg / mL, and the culture conditions were: 37°C, 5% CO 2cultured in a cell culture incubator. Monitor cell growth status by MTT method: remove the medium, wash with PBS buffer 3 times, add 20 μL of MTT reagent to each duplicate well, continue to stand in the cell culture incubator for 4 hours, remove the medium again, wash with PBS After washing with buffer solution for 3 times, 150 μL of DMSO reagent was added to each duplicate well, and the o...

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Abstract

The invention provides a method for enhancing the differentiation of mesenchymal stem cells into nerve cells. The method comprises the following steps: separating sheep placental cells, then breakingthe cells by using a repeated freeze-thaw process and acquiring sheep placenta extract; adding an appropriate amount of the sheep placenta extract into a stem cell culture solution; and adding a brain-derived neurotrophic factor in the optimal period of stem cell culture so as to obtain differentiated nerve cells. The method of the invention has the advantages that operation process is simple andis easy to learn, and at the same time, the method overcomes problems in proliferation and directed differentiation of bone marrow mesenchymal stem cells into nerve cells.

Description

technical field [0001] The invention belongs to the field of cell culture and enhanced differentiation of stem cells, in particular to a method for enhancing the differentiation of mesenchymal stem cells into nerve cells. Background technique [0002] Nervous system degenerative diseases belong to a class of progressive central nervous system diseases in clinical practice, which are mainly caused by the degeneration of systemic special nerve cell subgroups. Common types of diseases include Parkinson's disease, hereditary chorea, Alzheimer's Alzheimer's disease, etc., whose common feature is the loss or dysfunction of nerve cells, and stem cells have great potential in the treatment of these diseases. [0003] Under specific induction conditions in vivo or in vitro, mesenchymal stem cells can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, endothelial and other tissue cells. With multi-directional differentiation potential, me...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12N5/0775
CPCC12N5/0619C12N2500/84C12N2501/13C12N2506/1353
Inventor 华子昂
Owner 华子昂
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