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Preparation method of neural stem cells and glutamatergic neurons in cerebral cortex

A technology of nerve cells and nerves, applied in the field of preparation of cerebral cortex neural stem cells and glutamatergic neurons, can solve the problems of time-consuming, difficult to obtain high-purity neural stem cells and nerve cells, insufficient, etc., to reduce the risk of pollution , low cost, and the effect of promoting differentiation

Active Publication Date: 2020-02-07
广州瑞臻再生医学科技有限公司
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Problems solved by technology

In the field of stem cell research, these methods are notoriously inadequate, time-consuming (up to 4 weeks for neural induction with mouse stromal cells), and generate heterogeneous cell populations, including mesoderm, endoderm, and embryonic ectoderm , it is difficult to obtain high-purity functional neural stem cells and nerve cells for clinical cell therapy

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  • Preparation method of neural stem cells and glutamatergic neurons in cerebral cortex
  • Preparation method of neural stem cells and glutamatergic neurons in cerebral cortex
  • Preparation method of neural stem cells and glutamatergic neurons in cerebral cortex

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Embodiment 6

[0042] The present embodiment provides a method for preparing cerebral cortex neural stem cells and glutamatergic neurons. This embodiment adopts the formulation of the neural induction medium and the neural cell culture medium in Example 5, which specifically includes the following steps:

[0043] (1) Culture of human induced pluripotent stem cells (iPSCs): iPSCs were cultured with TeSR under the condition of using StemAdhere basement membrane matrix TM 2 or E8 and other serum-free animal-derived cell culture medium. Cells were passaged every 7 days using ReLeSR (1:6). Undifferentiated pluripotent stem cells usually express SSEA3, SSEA4, Tra-1-60, Tra-1-81, Oct4, and NANOG, which can be detected and identified by immunofluorescence staining and RT-PCR (eg figure 1 ),Depend on figure 1 It can be seen that the markers of iPSC cells (OCT4, SSEA4, SOX2, Tra-1-60, NANOG and Tra-1-81) are all expressed, indicating that they are in an undifferentiated state and have the function o...

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Abstract

The invention provides a preparation method of neural stem cells and glutamatergic neurons in cerebral cortex, and relates to an iPSC complete neural induction culture medium. The iPSC complete neuralinduction culture medium comprises the following components: a cell basic culture medium, a cell culture additive, neural development factors and an inhibitor. The invention provides the iPSC complete neural induction culture medium, an iPSC nerve cell culture medium and a culture method. The culture medium includes a serum-free basic medium, a cell additive, neural development factors and an inhibitor. The use of the serum-free basic medium can avoid the introduction of exogenous substances and reduce the risk of pollution. Meanwhile, a variety of nutrients, such as the cell additive and theneural development factors, can enable iPSC cells to be stimulated by signals so that iPSC cells can be activated to promote differentiation. The cell culture method provided by the invention is simple, efficient, low in cost, and can induce high-quality neuron cells in a short period.

Description

technical field [0001] The invention belongs to the technical field of cell differentiation, and in particular relates to a preparation method of cerebral cortex neural stem cells and glutamatergic neurons. Background technique [0002] It has been twelve years since Shinya Yamanaka and his team established the first human induced pluripotent stem cell line (hiPSC) in 2007, but because little is known about the molecular mechanisms that control their differentiation, the practical applications of these cells have been limited. Limited, specifically, the differentiation step from multifunctional hiPSCs to neuroectodermal cells (also known as neural induction) is poorly understood. [0003] So far, since the mechanism of neural induction is still unclear, the directed and complete differentiation of embryonic stem cells into neural tissue to reflect the embryogenesis process has not been realized. The generation of neural cells from hiPSCs is entirely based on empirical metho...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12N5/0797
CPCC12N5/0619C12N5/0623C12N2500/32C12N2500/90C12N2501/105C12N2501/11C12N2501/13C12N2501/155C12N2501/41C12N2501/727C12N2506/45
Inventor 张凌何翠敏
Owner 广州瑞臻再生医学科技有限公司
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