Method for reprogramming spinal marrow astrocytes into motor neurons through induction

Abstract

The invention provides a method for reprogramming spinal marrow astrocytes into motor neurons through induction. The method comprises steps as follows: (1) primary rat astrocytes are subjected to pure culture; (2) 7 small molecule drugs including SB431542, LDN-193189, RA, bFGF, Purmorphamine, Forskolin and VPA are selected according to substances required in the stages of neuron induction and motor neuron formation and development, and the astrocytes are induced and reprogrammed in vitro through the small molecule drugs; (3) the reprogramming and differentiation conditions of the astrocytes to the motor neurons are analyzed through observation of morphologic change of cells and methods of chemical technology analysis of immunofluorescent cells, real-time fluorescent quantitative PCR analysis and statistical analysis. The rat astrocytes can be successfully induced and reprogrammed into the motor neutrons in vitro through small molecular drug composition, and the transdifferentiation exceeds 75%; in-situ direct reprogramming of the astrocytes to the neurons is a possibly comparatively feasible thought in SCI (spinal cord injury) repairing and functional reconstruction treatment.

Description

technical field [0001] The invention belongs to the field of medical technology, and in particular relates to a method for inducing reprogramming of spinal cord astrocytes into motor neurons. Background technique [0002] The clinical symptoms of spinal cord injury (SCI) range from slight sensory and motor impairments to complete paralysis of limbs, mainly depending on the degree and scope of the injury. Astrocytes play a decisive role in the nutrition, support and regulation of neurons under normal physiological conditions. However, once the spinal cord is injured, astrocytes will quickly activate into reactive astrocytes. And continue to proliferate, forming a glial scar around the injury site. Although glial scar can control the spread of inflammation and protect surviving neurons and other cells, its formation also constitutes a physical and chemical barrier, which seriously hinders the regeneration of axons, and some studies have also found that some reactive glue Pla...

AI Technical Summary

Problems solved by technology

However, when it is applied to the repair of spinal cord injuries, researchers still find that there are many application limitations

First, transcription factors commonly used for reprogramming are highly expressed in various types of tumors, which will make iPSCs have higher tumorigenicity; second, about 2 weeks after spinal cord injury is the best period for nerve repair, while The entire induction and differentiation time of iPSCs is too long, and the differentiation and repair cannot be completed within the optimal repair period; moreover, iPSCs cannot guarantee the directed and efficient differentiation into neurons, especially motor neurons, in the complex spinal cord environment.

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