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

In vitro generation of gabaergic neurons from embryonic stem cells and their use in the treatment of neurological disorders

A technology of embryonic stem cells and neurons, applied in animal cells, nervous system cells, vertebrate cells, etc., can solve the problem of not being able to produce high-yield GABA neurons

Inactive Publication Date: 2006-11-29
RELIANCE LIFE SCI PVT
View PDF12 Cites 5 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

A method for obtaining cells producing γ-aminobutyric acid from mouse embryonic stem cells has also been reported (Hancock et al., 2000, Biochem.Biophys.Res.Commun.271 (2): 418-21, Westmoreland et al., 2001 , Biochem.Biophys.Res.Commun.284(3):674-80; U.S.Publication No.2003 / 0036195A1, each of which is specifically incorporated herein by reference), but none of these methods can produce high yields of GABA neurons

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
  • In vitro generation of gabaergic neurons from embryonic stem cells and their use in the treatment of neurological disorders
  • In vitro generation of gabaergic neurons from embryonic stem cells and their use in the treatment of neurological disorders
  • In vitro generation of gabaergic neurons from embryonic stem cells and their use in the treatment of neurological disorders

Examples

Experimental program
Comparison scheme
Effect test

preparation example Construction

[0077] Preparation of neuroblasts

[0078] Isolated pluripotent stem cells can be expanded and then transferred to culture conditions that differentiate into neuroblasts. Pluripotent stem cells were cultured according to the differentiation protocol described here to enter the neural cell differentiation pathway. Pluripotent stem cells are cultured on a suitable substrate in a differentiation nutrient medium containing differentiating agents such as soluble factors and growth factors. Suitable substrates include, but are not limited to, solid surfaces coated with positive charges, such as poly-L-lysine or polyornithine, substrates coated with extracellular matrix components, such as fibronectin, laminin, platelet-derived Growth factor (PDGF), epidermal growth factor (EGF), collagen V, human amnion or recombinant cell basement membrane (Matrigel  ), or a combination of them. A preferred differentiation nutrient medium is capable of maintaining the proliferation, differentia...

example 1

[0122] The following example demonstrates the in vitro production of functional GABAergic neurons from mouse embryonic stem cells. figure 1 produced GABAergic neurons from murine ES cells, while figure 2 are different steps in differentiating murine ES cells into terminally differentiated neurons.

[0123] 1) Culture and expansion of mouse embryonic stem cells:

[0124] The mouse ES cells used in this experiment were isolated from the inner cell mass of mouse blastocysts, and the techniques used are well known to those skilled in the art. Murine ES cells were of J-1 origin (obtained from National Institute of Dental and Craniofacial Research, National Institute of Health, Bathesda, Maryland, USA) and passaged for 14 passages. These cells were cultured on mitomycin-C treated mouse embryonic fibroblast feeder cells, which arrests mitosis in ES cell culture medium. Murine ES cells are cultured in ES cell culture medium to expand the number of undifferentiated cells. ES cells...

example 2

[0152] In the following example, the present applicants used an in vitro transplantation model to study the potency, viability and function of GABAergic neurons derived from murine embryonic stem cells. figure 1 It is an in vitro transplantation model.

[0153] The survival and function of GABAergic neurons derived from murine ES cells were studied by ex vivo transplantation of GABAergic neurons and hippocampal cells of adult rats. Hippocampal cells were first isolated from isolated cells of adult mouse brain hippocampus, and then dispersed with 0.05% trypsin-EDTA, followed by Neurobasal supplemented with B27 (20 μg / ml) and N2 (10 μg / ml). Cells were cultured on -A medium for 1 week. Adult hippocampal brain cells were cultured on 100 mm tissue culture plates coated with poly-L-ornithine and laminin or gelatin. After 1 week, GABAergic neurons derived from murine ES cells and harvested after 12 days of differentiation were added to adult hippocampal brain cells, and the two cel...

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

The present disclosure is directed to improved methods for efficiently producing neuroprogenitor cells and differentiated neural cells such as GABAergic neurons from pluripotent stem cells, for example embryonic stem cells. Using the disclosed methods, cell populations containing a high proportion of GABAergic neurons have been isolated. The neuroprogenitor cells and terminally differentiated cells of the present disclosure can be generated in large quantities, and therefore may serve as an excellent source for cell replacement therapy in neurodegenerative disorders and neuronal diseases such as stroke, ischemia, epilepsy, and Huntington's disease.

Description

technical field [0001] The present invention relates to an improved method for producing terminally differentiated neuronal cells, such as GABAergic neurons, from pluripotent stem cells, such as mouse or human embryonic stem cells. The GABAergic neurons involved in the present invention can provide excellent cell replacement therapy for neurodegenerative diseases and neuronal diseases, such as stroke, ischemia, Parkinson's disease, Alzheimer's disease, epilepsy and Huntington's disease. source of cells. Background technique [0002] Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter in the central nervous system (CNS), which is widely distributed in brain tissue and expressed in interneurons that regulate local circulation. GABAergic neurons capable of producing GABA are the major inhibitory neurons in the mammalian central nervous system, where 60-75% of synapses are GABAergic neurons (Schwartz, R.D., 1988, Biochem. Pharmacol. 37:3369-75). GABAergic ...

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): C12N5/00C12N5/06C12N5/079C12N5/0793
Inventor 菲尔多斯·阿拉姆·肯萨蒂什·马哈德奥罗·托泰
Owner RELIANCE LIFE SCI PVT
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