Neurons directly induced from human skin cells and preparation method for neurons

Abstract

The invention belongs to the field of biomedicine and relates to neurons directly induced from human skin cells and a preparation method for the neurons. By the method, induced human neurons which can perform stable passage are directly prepared by importing complementary deoxyribonucleic acid (cDNA) containing reprogramming transcription factors, namely Sox2, Ascl1 and Myt1l into isolated adult skin cells, screening cells which express a human neuron marker and cloning the cells which express the human neuron marker. The invention is characterized in that (1) the neurons express human neuron-specific marker protein in vitro; and (2) the neurons can survive in vivo and exert functions. The human neurons with the functions are obtained without the process of multipotential stem cells, so that the technology is relatively simple and effective; a heterologous feeder layer is avoided, so that the neutrons have a good clinical application prospect; and the report related to the tumorigenesis is not known about the adopted transcription factors, so the neurons have relatively high biosafety.

Description

technical field [0001] The invention belongs to the field of biomedicine and relates to a method for inducing neuron cells, in particular to a method for directly inducing neuron cells from human skin cells. Background technique [0002] Nervous system diseases such as brain injury, spinal cord injury, stroke, Parkinson's disease, neuronal amyotrophic lateral sclerosis, Alzheimer's disease, etc. have great social harm and difficult treatment, and have long been difficult problems in the medical field. Studies have shown that using human neuron cells to study the mechanism of nerve injury, screen new drugs that interfere with nerve function, and use them for transplantation to repair damaged nerve function is an effective and promising method; however, the source of functional nerve cells is limited. Commonly used embryonic stem cells (embryonic stem cells, ES cells) differentiated to obtain human neurons (Wichterle H, Lieberam I, Porter JA, et al. Directed differentiation of...

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Problems solved by technology

[0005] However, the above-mentioned technical methods still have the following defects: 1) so far, human skin cells cannot be induced by using the combination of transcription factors that induce mice; 2) in animal experiments, human inducible neurons can not be obtained Through transgenic technology, the reporter gene that marks the change of cell type is integrated into the mouse genome to make a transgenic mouse. After the skin cells are transformed into nerve cells, the required nerve cells can be selected by fluorescence or antibiotic resistance screening. , but because transgenic technology cannot be directly applied to humans, this cell screening method cannot be applied to the preparation of inducible human neuron cells; 3) It is generally believed that to fully evaluate the characteristics and functions of a new class of cells, a complete and in vivo experimental data, but the aforementioned studies only carried out in vitro experiments when evaluating mouse induced neuron cells, and did not transplant the obtained functional cells into the body for analysis and identification, thus lacking a complete understanding of the characteristics of this new type of cells

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