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Method for inducing human or animal somatic cells to neural stem cells

A neural stem cell and cell induction technology, 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 safe application

Active Publication Date: 2017-12-05
ZHONGSHAN OPHTHALMIC CENT SUN YAT SEN UNIV
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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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  • Method for inducing human or animal somatic cells to neural stem cells
  • Method for inducing human or animal somatic cells to neural stem cells
  • Method for inducing human or animal somatic cells to neural stem cells

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Embodiment Construction

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

[0048] The specific process and details of the present invention will be described in detail below by taking "Ptf1a lentivirus transdifferentiates 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 the lentiviral vector (Tet-O-FUW) containing the tet-on regulatory sequence. The purpose is to co-transfect with RTTA virus in the future, and Dox (Doxycyclin) can be added to control the expression of Ptf1a in somatic cells. After the correct cloning was verified by DNA sequencing, the plasmids for transfection were prepared.

[0051] (2) Preparation of lentivirus

[0052] 1) 24 hours before plasmid transfection, spread 4.0x10 cells in a 10cm cell culture dish 6 HEK293T cells were cultured by adding 6ml MEF medium.

[0053] 2) Before transfection, remove the...

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Abstract

The invention discloses a method for inducing human or animal somatic cells to neural stem cells and purpose of neural stem cells obtained by the method. By overexpression of transcription factor Ptf1a in separated human or animal somatic cells and culture in a culture medium containing EGF and bFGF, somatic cells can be induced to neural stem cells (iNSCs). The iNSCs obtained by the method has self-renewing capability, and can stably passage for 50 generations and above and maintain original cell characteristics. By culture in different in vitro differentiation mediums, the iNSCs can be efficiently differentiated to neurons, astrocytes and oligodendrocytes. By injecting the iNSCs in animal bodies, the injected cells can be integrated into nervous tissues of retina, cerebrum, etc. the method of the invention is simpler, safer and more efficient than previous methods. The iNSCs obtained by the method and further differentiated cells can be applied to cell models of nerve diseases, drug screening, stem cell treatment and the like.

Description

technical field [0001] The present invention relates to the field of biotechnology such as stem cells, cell tissue regeneration, and cell replacement therapy, and mainly uses transcription factors to induce transdifferentiation of human or animal somatic cells into neural stem cells in vitro, and provides a new way to obtain induced neural stem cells Compared with other methods, it is simpler, safer and more efficient, and has great applications in basic research of neurodegenerative diseases, drug screening and stem cell therapy. 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 increasing, which brings serious harm to individuals, families and society. Health threats and heavy financial burdens. The common feature of these diseases is the gradual death of local nerve cells...

Claims

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Application Information

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Patent Type & Authority Applications(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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