Method for in vitro amplifying, and in 3D solid culturing nerve stem

A neural stem cell, three-dimensional technology, applied in the field of neural stem cell culture, can solve the problems that are difficult to meet the clinical application, low survival rate, and small number of neural stem cells, so as to facilitate the proliferation and division, expand the culture area, improve micro environmental effects

Inactive Publication Date: 2007-12-26
CHINA JAPAN FRIENDSHIP HOSPITAL
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Problems solved by technology

[0002] For a long time, the technology of separating and purifying neural stem cells from embryos or fetal brains by using neural stem cell culture medium and cell cloning technology has been basically perfecte

Method used

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  • Method for in vitro amplifying, and in 3D solid culturing nerve stem

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Embodiment Construction

[0014] The method for in vitro expansion of neural stem cells in three-dimensional culture of the present invention comprises:

[0015] Choose a porous microcarrier with a three-dimensional environment, for example: choose Cytopore from Amersham Biosciences, Sweden TM 2 microcarriers;

[0016] Pretreat the microcarriers, such as: put 1g of microcarriers into 100ml of PBS buffer, soak for 30 minutes at room temperature, and invert the microcarriers from time to time to make the microcarriers fully contact with the buffer, and then heat the microcarriers at about 120°C Under the temperature and the pressure of about 0.11 MPa, the above-mentioned microcarriers were sterilized for 20 minutes, and then the above-mentioned microcarriers were washed twice with PBS buffer solution with a pH value of about 7.4, and after being washed once with serum-free culture medium, the above-mentioned microcarriers were The carrier is placed in a refrigerator at about 4°C overnight;

[0017] The...

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Abstract

This invention relates to a method for amplifying neural stem cells in vitro by 3-dimensional culture. The method comprises: selecting microcarrier with 3-dimensional environment, pre-treating, coating the microcarrier with 40-60 ng/mL alkaline fibroblast growth factor, 40-60 ng/mL epidermal growth factor, and B27 DMEM/F12 neural stem cell serum-free culture medium, adding 1X105-1X106 neural stem cells into the culture bottle, taking out the microcarrier grown with neural stem cells, removing the microcarrier, and rinsing cells to obtain neural stem cells. The porous microcarrier can enlarge the culture area. The alkaline fibroblast growth factor and epidermal growth factor can promote cell multiple fission and improve cell microenvironment, which is advantageous for multiple fission of neural stem cells.

Description

technical field [0001] The invention relates to a method for culturing neural stem cells, in particular to a method for in vitro expansion of neural stem cells in three-dimensional culture. Background technique [0002] For a long time, the technology of separating and purifying neural stem cells from embryos or fetal brains by using neural stem cell culture medium and cell cloning technology has been basically perfected, but the problem of long-term culture of neural stem cells in vitro is still unsolved. The number of neural stem cells is small, and the survival rate is low, which is difficult to meet the clinical application. Contents of the invention [0003] The purpose of the present invention is to provide a method for in vitro expansion of neural stem cells in three-dimensional culture. [0004] In order to achieve the purpose of this invention, the present invention provides a method for in vitro expansion of neural stem cells in three-dimensional culture, which ...

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Application Information

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IPC IPC(8): C12N5/06C12N5/08C12N5/0797
Inventor 蔡哲潘琳张岚舒峻房青
Owner CHINA JAPAN FRIENDSHIP HOSPITAL
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