High-low density anchorage-dependent cell culturing method of old embryonic age neural stem cells

Abstract

The invention provides a high-low density anchorage-dependent cell culturing method of old embryonic age neural stem cells. The method comprises the steps of: separation and culture of the cortex neural stem cells, culture of the neural stem cells by the high-low density anchorage-dependent cell culturing method, subculture of the neural stem cells, monoplast cloning of the neural stem cells, differentiation culture of the neural stem cells and detection by an immumofluorescence method. According to the method provided by the invention, self-renewal neural stem cells with the differentiative potential are separated and cultured from an embryonic human brain cortex which is 13 weeks old by serum-free culture and monoplast clone technology. A plenty of neural stem cells are cultured by the high-low density anchorage-dependent cell culturing method, wherein the density decreases gradually. The cells have the continuous cloning capacity, can be subcultured, and express neural nidogen antigen. Differentiated cells express the specific antigens of neuron, gliocyte and oligodendroglia cell, so that the problem that embryo neural stem cells are hard to culture is solved.

Description

technical field [0001] The invention relates to a method for culturing embryonic neural stem cells in the field of biotechnology, in particular to a method for culturing high-low density adherent cells of large embryonic neural stem cells. Background technique [0002] The discovery of neural stem cells is a major breakthrough in the field of neuroscience, which has opened up broad prospects for research fields such as neural development and neural tissue transplantation. Neural stem cells are cells with self-renewal and extensive differentiation potential. They are in a non-terminal state of differentiation. They can divide symmetrically or asymmetrically into new stem cells or daughter cells with gradually decreasing differentiation potential, and finally produce the central nervous system. The three main types of cells are neurons, astrocytes, and oligodendrocytes. It is generally believed that neural stem cells mainly exist in the striatum, cerebellar hemispheres, and v...

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problem that embryonic neural stem cells are difficult to cultivate, the present invention uses serum-free culture and single cell cloning technology to isolate and cultivate neural stem cells with self-renewal and multi-differentiation potentials from the human embryonic cerebral cortex at 13 weeks. The gradually decreasing high-low density adherent cell culture method can form a large number of neural stem cells after 3 weeks. The cells have continuous cloning ability, can be conveyed and cultured, and express neural nestin antigen; the differentiated cells express neuronal cells, glue oligodendrocyte specific antigen

Images