Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Method for efficiently and rapidly reprogramming somatic cells into neural stem cells and application thereof

A neural stem cell and reprogramming technology, applied in the field of efficient and rapid reprogramming of somatic cells into neural stem cells, can solve the problems of low efficiency of neural stem cell clone formation, low efficiency of somatic cell transformation, and long period of somatic cell transdifferentiation. Beneficial for clinical application, high reprogramming efficiency, avoiding the effect of mutation or integration

Active Publication Date: 2021-04-06
BINZHOU MEDICAL COLLEGE
View PDF4 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] The technical problem to be solved by the present invention is mainly to overcome the problems of low reprogramming somatic cell transformation efficiency, low neural stem cell clone formation efficiency and somatic cell transdifferentiation in the prior art. Long cycle and other defects, so as to provide a method for inducing neural stem cells that can use their own body cells to reprogram quickly, with high transdifferentiation efficiency and short transdifferentiation time

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Method for efficiently and rapidly reprogramming somatic cells into neural stem cells and application thereof
  • Method for efficiently and rapidly reprogramming somatic cells into neural stem cells and application thereof
  • Method for efficiently and rapidly reprogramming somatic cells into neural stem cells and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0061] The principles and features of the present invention are described below in conjunction with examples, which are only used to explain the present invention and are not intended to limit the scope of the present invention.

[0062] Step 1: Isolation and culture of human foreskin fibroblasts

[0063] (1) Get the fresh foreskin tissue just removed from the operation and rinse it with PBS for 3 times to remove blood stains;

[0064] (2) Use tweezers and scissors to remove the attached subcutaneous fat tissue, capsule connective tissue and necrotic tissue, rinse with PBS 3 times, and cut into about 1mm 3 small pieces;

[0065] (3) Transfer the tissue into a centrifuge tube, add a little trypsin, and incubate in a 37°C incubator for 30 minutes;

[0066] (4) Centrifuge at 1000rpm for 5min, discard the supernatant, add 1mmg / ml collagenase type IV, and incubate in a 37°C incubator for 30min;

[0067] (5) Add hPFF medium to stop digestion, centrifuge at 1000rpm for 5min, disca...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

One of the purposes of the invention is to provide a method for efficiently and rapidly reprogramming somatic cells into neural stem cells. The method comprises the following steps: separating, extracting and amplifying somatic cells; transfecting a gene knock-down vector or a gene knock-out vector; carrying out multiplication culture on the neural stem cells; the gene knock-down vector or the gene knock-out vector comprises shRNA, siRNA, a CRISPR gene editing vector or a related microRNA inhibitor for blocking let-7 genes. According to the present invention, the expression of one microRNA-let-7b is knocked down or knocked out without introducing any exogenous gene, and the neural stem cells are successfully induced from fibroblasts, such that the sufficient amount of cells required by transplantation can be generated through amplification within a short time, and the cells are genetically stable and can be differentiated to form neurons, astrocytes and oligodendrocytes; more than 20 generations of neural stem cells can be amplified, and no induced tumor is formed after the neural stem cells are transplanted to mice.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a method for efficiently and rapidly reprogramming somatic cells into neural stem cells and its application. Background technique [0002] The development of stem cell technology has brought hope for the treatment of many neurological diseases, including neurodegenerative diseases such as Alzheimer's disease and Parkinson's disease, neurological damage diseases such as spinal cord injury, and amyotrophic lateral sclerosis syndrome, autism, etc. Studies have shown that the use of neurons differentiated from embryonic stem cells has a certain effect in treating diseases such as Alzheimer's disease, Parkinson's disease, and spinal cord injury. However, the source of embryonic stem cells is ethically controversial, and its tumorigenicity and immune rejection limits its application. . [0003] Induced pluripotent stem cells (iPSCs) formed by reprogramming of somatic cells ca...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C12N5/10C12N5/0797C12N15/113A61K35/30A61P25/28A61P25/16A61P25/00A61P25/14A61P9/10
CPCC12N5/0623C12N15/113A61K35/30A61P25/28A61P25/16A61P25/00A61P25/14A61P9/10C12N2506/1307C12N2501/115C12N2501/11C12N2500/38C12N2500/30C12N2533/32C12N2533/52C12N2533/54
Inventor 王跃嗣
Owner BINZHOU MEDICAL COLLEGE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com