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Method for inducing differentiation of neural stem cells

A neural stem cell, directional differentiation technology, applied in biochemical equipment and methods, nervous system diseases, microorganisms, etc., can solve the problems of easy loss of stem cell characteristics, limited theoretical research and clinical application value, and high apoptosis rate. The effect of clinical application prospects

Inactive Publication Date: 2010-05-12
王彦哲
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Problems solved by technology

However, the technique of inducing bone marrow stromal cells to differentiate into neural stem cells is very difficult, and there are many uncertain factors. At present, the commonly used methods such as retinoic acid and nerve growth factor are used to induce bone marrow stromal cells to differentiate into neural cells. It is very easy to lose stem cell characteristics and cell apoptosis. The rate is extremely high, which greatly limits the value of theoretical research and clinical application

Method used

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Embodiment 1

[0013] 1. Establishment of adult stem cell lines. Firstly, separate adult stem cells from a variety of embryonic tissues. After purification, they are cultured for 20-40 generations under serum-free conditions in vitro to establish hematopoietic cells, neural stem cells, liver cells, skin Adult stem cell lines such as stem cells and muscle stem cells. Cells are required to grow in good condition, passage every 3 to 4 times, without contamination by bacteria, mycoplasma, etc.; passaged cells need to be tested by cytogenetics, clone formation experiment, nude mouse tumorigenicity experiment, etc., to confirm that they are cells in normal growth state .

[0014] 2. Directional induction of MSCS: After changing the culture medium for 2 days, change the medium and add 0.01-100mMOL / L monosialotetrahexose ganglion lipid (GMI) to induce 1, 3, 5, 7, 14, 28 days; In the control group, only Neurobasal medium containing 2% B27 was cultured for the same time.

[0015] 3. Observation of MSCS a...

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Abstract

The invention provides a method for inducing differentiation of neural stem cells, belongs to the field of biotechnology, and in particular relates to a method for inducing differentiation of neural stem cells by an adult stem cell line. The invention provides a method for inducing the differentiation of the neural stem cells, which comprises the following steps of: 1, establishment of the adult stem cell line, namely separating and purifying animal embryo tissues under an aseptic condition to form adult stem cells, and generating different stem cell lines through tens to thousands of subculture; 2, MSCS induction, namely replacing culture solution after 2 days and adding 0.01 to 100Mmol / l monostalotetrahexosylgangliside to induce respectively; 3, morphological observation of SCS and the induced cells; and 4, immunocyte chemical dyeing, namely performing cell immunofluorescent dyeing on the induced cells with a nidogen, an NSE and a GFAP antibody respectively.

Description

Technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for inducing the directional differentiation of neural stem cells, in particular to a method for inducing the directional differentiation of neural stem cells by using adult stem cell lines. Background technique [0002] Neural stem cells have broad application prospects in the treatment of nerve tissue damage and repair. Neural stem cells are mainly derived from embryonic tissue and adult tissue cells. Adult stem cells include hematopoietic stem cells, neural stem cells, liver stem cells, skin stem cells, muscle stem cells, and so on. [0003] At present, research reports on the induction and differentiation of stem cells derived from adult non-neural tissues into neural stem cells have gradually increased, mainly focusing on bone marrow, fat, skin and other tissues, among which adult neural stem cells derived from bone marrow have been studied more. The main method is to extract a...

Claims

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Application Information

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IPC IPC(8): C12N5/06A61P25/16A61P9/10A61P25/00A61P25/28
Inventor 王彦哲
Owner 王彦哲
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