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Preparation method and application of induction type neural stem cells

A neural stem cell, inducible technology, applied in nervous system cells, botanical equipment and methods, biochemical equipment and methods, etc., can solve the problems of iNs cells being unable to undergo cell division, difficult to survive in vivo, and restricting clinical applications.

Active Publication Date: 2012-12-19
INST OF ZOOLOGY CHINESE ACAD OF SCI
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Problems solved by technology

Although iPSCs cells can continuously produce neural precursor cells, the tumorigenicity problem after transplantation restricts the clinical application of this kind of pluripotent stem cells (Wer nig, M., et al., Neurons derived from reprogrammed fibroblasts functionally integrate into the fetal brain and improve symptoms of rats with Parkinson's disease. Proc Natl Acad Sci U S A, 2008.105(15): p.5856-61.)
On the contrary, iNs cells cannot undergo cell division at all, so it is difficult to survive in vivo after transplantation back to the diseased area
In addition, the iNs cells obtained in previous studies can only be a type of peripheral neurons, and other types of neurons have not been reported by direct induction.

Method used

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  • Preparation method and application of induction type neural stem cells
  • Preparation method and application of induction type neural stem cells
  • Preparation method and application of induction type neural stem cells

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Experimental program
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Embodiment 1

[0036] Isolation of Sertoli cells:

[0037] Utilize an improved method (Richardson, L.L., H.K.Kleinman, and M.Dym, Basement membrane gene expression by Sertoli and peritubular myoid cells in vitro in the rat. Biol Reprod, 1995.52 (2): p.320-30.) can Separation of Sertoli cells of high purity. Specific steps are as follows:

[0038] The testes of the 5-day-old mice were removed. Remove the albuginea of ​​the testis. Digest in three steps then: (one) 0.1% (mass percentage) collagenase IV, digest for 20 minutes at 37 ℃; (2) simultaneously use 0.1% (mass percentage) collagenase IV and 0.1% (mass percentage) hyaluronic acid Enzymes, digested at 37°C for 10 minutes, washed with PBS after digestion; (3) 0.1% (mass percentage) collagenase IV, 0.1% (mass percentage) hyaluronidase, 0.25% (mass percentage) trypsin, 0.04% (mass percentage) DNase I mixture, digested at 37°C for 20 minutes, terminated digestion with fetal bovine serum (FBS, Invitrogen), filtered cells with 200-mesh cell...

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Abstract

The invention provides a preparation method and application of induction type neural stem cells. The preparation method of the induction type neural stem cells comprises the steps of: carrying out overexpression of exogenous factors Ascl1, Neurog2, Pax6, Hes1, Id1, Brn2, c-Myc and Klf4 in sustentacular cells of testis, and culturing in a base culture fluid containing cell growth factors to obtain the induction type neural stem cells. According to the preparation method of the induction type neural stem cells, sustentacular cells of testis derived from the mesoblast are reprogrammed to be neural stem cells derived from the ectoderm, the neural stem cells obtained in the reprogramming manner are proved to have physiological functions in vivo or in vitro, and the cells are safer than pluripotent stem cells for transplantation.

Description

technical field [0001] The invention relates to a preparation method and application of inducible neural stem cells, in particular to a rapid and efficient preparation method and application of inducible neural stem cells that enable cells to directly transdifferentiate cells across the germ layer without going through a pluripotent state. Background technique [0002] People are paying more and more attention to the pathogenesis and treatment of neurological diseases year by year. In today's society, science and technology are advancing rapidly, and new drugs are being developed in an endless stream. However, it is undeniable that the number of patients with neurological diseases is increasing day by day worldwide. Neurodegenerative diseases such as Parkinson's disease and Alzheimer's disease have seriously reduced the quality of life of patients themselves and increased the burden of life on patients' families. Therefore, how to fundamentally treat neurological diseases, ...

Claims

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Application Information

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IPC IPC(8): C12N5/10C12N5/0797C12N15/867C12N5/071C12Q1/02A61L27/38
Inventor 周琪盛超郑钦元王柳李伟赵小阳
Owner INST OF ZOOLOGY CHINESE ACAD OF SCI
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