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A method for inducing human or animal somatic cells into neural stem cells

A neural stem cell and cell induction technology, which is applied in the field of inducing human or animal somatic cells into neural stem cells, can solve the problems of safety and hidden dangers of transcription factors, and achieve the effect of application safety

Active Publication Date: 2021-02-09
ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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  • Description
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Problems solved by technology

[0016] The purpose of the present invention is to provide a method for inducing human and animal somatic cells into neural stem cells, solve the problem of potential safety hazards in transcription factors, and diversify the methods of transdifferentiation

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  • A method for inducing human or animal somatic cells into neural stem cells
  • A method for inducing human or animal somatic cells into neural stem cells
  • A method for inducing human or animal somatic cells into neural stem cells

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Embodiment Construction

[0047]The following examples are a further detailed description of the present invention.

[0048]The specific process and details of the present invention will be described in detail below with "Ptf1a lentivirus transdifferentiating fibroblasts into iNSCs" as an example.

[0049](1) Construction of lentiviral vector (Tet-O-FUW-Ptf1a)

[0050]The mouse Ptf1a (NM_018809.2, 972bp) coding sequence was cloned into a lentiviral vector (Tet-O-FUW) containing tet-on regulatory sequence. The purpose is to co-transfect with RTTA virus in the future and add Dox (Doxycyclin) to control the expression of Ptf1a in somatic cells. After the correct clone was verified by DNA sequencing, the plasmid for transfection was prepared.

[0051](2) Preparation of lentivirus

[0052]1) 24 hours before plasmid transfection, spread 4.0x10 in a 10cm cell culture dish6HEK293T cells were cultured by adding 6ml MEF medium.

[0053]2) Before transfection, remove the original medium and replace it with 9ml fresh pre-warmed MEF mediu...

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Abstract

The invention discloses a method for inducing human or animal somatic cells into neural stem cells and the use of the neural stem cells obtained by the method. Somatic cells can be induced into neural stem cells (induced neural stem cells, iNSCs) by overexpressing the transcription factor Ptf1a in isolated human or animal somatic cells and culturing them in a medium containing EGF and bFGF. The iNSCs obtained by this method have the ability of self-renewal, can be stably passaged for more than 50 passages, and maintain the original cell characteristics. Cultured in different in vitro differentiation media, iNSCs can efficiently differentiate into neurons, astrocytes, and oligodendrocytes. Injecting iNSCs in animals, the injected cells can be integrated into neural tissues such as retina and brain. The method used in the present invention is simpler, more efficient and safer than previous methods. The iNSCs obtained by the method of the present invention and the further differentiated cells thereof can be applied to cell models of neurological diseases, drug screening, stem cell therapy and the like.

Description

Technical field[0001]The present invention relates to the fields of biotechnology such as stem cells, cell tissue regeneration, cell replacement therapy, etc. It mainly uses transcription factors to induce and transdifferentiate human or animal somatic cells into neural stem cells in vitro, and provides a new way to induce neural stem cells. Compared with other methods, it is simpler, safer and more efficient, and has great applications in basic research, drug screening and stem cell treatment of neurodegenerative diseases.Background technique[0002]With the continuous progress of the global aging society, the incidence of neurodegenerative diseases represented by Alzheimer’s disease, Parkinson’s disease, age-related macular degeneration, retinitis pigmentosa, etc. is getting higher and higher, bringing serious consequences to individuals, families and society. Health threats and heavy economic burdens. The common feature of these diseases is the gradual death of local nerve cells, b...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/85C12N15/87C12N15/88C12N15/867C12N15/86C12N15/861C12N15/864C12N15/90C12N5/10
CPCC12N5/0623C12N15/85C12N15/86C12N15/87C12N15/88C12N15/90C12N2510/00C12N2710/10043C12N2740/15043C12N2750/14143C12N2760/18843
Inventor 向孟清晋康新邹珉肖冬长
Owner ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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