Method for reprogramming spinal marrow astrocytes into motor neurons through induction

A technology of astrocytes and motor neurons, applied in the medical field, can solve the problems of long induction and differentiation of iPSCs, inability to guarantee differentiated neurons, high tumorigenicity of iPSCs, etc.

Active Publication Date: 2019-09-27
NANTONG UNIVERSITY
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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 b

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  • Method for reprogramming spinal marrow astrocytes into motor neurons through induction
  • Method for reprogramming spinal marrow astrocytes into motor neurons through induction
  • Method for reprogramming spinal marrow astrocytes into motor neurons through induction

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Embodiment Construction

[0073] In order to make the technical problems, technical solutions and advantages to be solved by the present invention clearer, the following will describe in detail with reference to the drawings and specific embodiments.

[0074] A method for inducing reprogramming of spinal astrocytes into motor neurons, comprising the following steps:

[0075] (1) Purification and culture of primary rat astrocytes. Primary astrocytes from the spinal cord of neonatal rats were used as target cells to induce reprogramming, and were obtained by in vitro culture and microwave shock culture by differential attachment method Astrocytes with high purity; specifically include the following processes:

[0076] (1-1) The ultra-clean workbench is irradiated with ultraviolet rays for 30 minutes in advance, and the -20°C pre-cooled ice platform, sterilized surgical instruments, HBSS buffer, and plastic culture dishes are taken out and placed in the ultra-clean workbench, and the culture dishes are ad...

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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...

Claims

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Application Information

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IPC IPC(8): C12N5/0793C12N5/079
CPCC12N5/0619C12N2506/08C12N2501/115C12N2501/727
Inventor 王晓冬徐磊吴坚陈雪杨日云陈罡李奕陈颖潘静莹
Owner NANTONG UNIVERSITY
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