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Functional Assessment of Direct Reprogrammed Neurons In Vitro and In Vivo

Kidnapillai, Srisaiyini LU and Ottosson, Daniella Rylander LU (2021) In Methods in Molecular Biology 2352. p.183-199
Abstract

Direct reprogramming is an emerging research field where you can generate neurons from a somatic cell, such as a skin or glial cell by overexpressing neurogenic transcription factors. This technique allows fast generation of subtype-specific and functional neurons from both human and mouse cells. Despite the fact that neurons have been successfully generated both in vitro and in vivo, a more extensive analysis of the induced neurons including phenotypic functional identity or gradual maturity is still lacking. This is an important step for a further development of induced neurons towards cell therapy or disease modeling of neurological diseases. In this protocol, we describe a method for functional assessment of direct reprogrammed... (More)

Direct reprogramming is an emerging research field where you can generate neurons from a somatic cell, such as a skin or glial cell by overexpressing neurogenic transcription factors. This technique allows fast generation of subtype-specific and functional neurons from both human and mouse cells. Despite the fact that neurons have been successfully generated both in vitro and in vivo, a more extensive analysis of the induced neurons including phenotypic functional identity or gradual maturity is still lacking. This is an important step for a further development of induced neurons towards cell therapy or disease modeling of neurological diseases. In this protocol, we describe a method for functional assessment of direct reprogrammed neuronal cells both in vitro and in vivo. Using a synapsin-driven reporter, our protocol allows for a direct identification of the reprogrammed neurons that permits functional assessment using patch-clamp electrophysiology. For in vitro reprogramming we further provide an optimized coating condition that allows a long-term maturation of human induced neurons in vitro.

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Please use this url to cite or link to this publication:
author
and
organization
publishing date
type
Chapter in Book/Report/Conference proceeding
publication status
published
subject
keywords
AAV, Action potential, Direct conversion, Fibroblast, GFP reporter, Glia, iNs, Intracerebral injections, iPSC, Lentivirus, Postsynaptic activity, Regeneration, Stem cells, Striatum, Transdifferentiation
host publication
Neural Reprogramming: Methods and Protocols
series title
Methods in Molecular Biology
volume
2352
pages
183 - 199
publisher
Humana Press
external identifiers
  • scopus:85111805359
  • pmid:34324188
ISSN
1940-6029
1064-3745
ISBN
978-1-0716-1601-7
978-1-0716-1600-0
DOI
10.1007/978-1-0716-1601-7_13
language
English
LU publication?
yes
id
ed3b1ced-4c69-44e0-b58c-36e52881de09
date added to LUP
2021-12-10 09:29:58
date last changed
2024-06-01 21:46:34
@inbook{ed3b1ced-4c69-44e0-b58c-36e52881de09,
  abstract     = {{<p>Direct reprogramming is an emerging research field where you can generate neurons from a somatic cell, such as a skin or glial cell by overexpressing neurogenic transcription factors. This technique allows fast generation of subtype-specific and functional neurons from both human and mouse cells. Despite the fact that neurons have been successfully generated both in vitro and in vivo, a more extensive analysis of the induced neurons including phenotypic functional identity or gradual maturity is still lacking. This is an important step for a further development of induced neurons towards cell therapy or disease modeling of neurological diseases. In this protocol, we describe a method for functional assessment of direct reprogrammed neuronal cells both in vitro and in vivo. Using a synapsin-driven reporter, our protocol allows for a direct identification of the reprogrammed neurons that permits functional assessment using patch-clamp electrophysiology. For in vitro reprogramming we further provide an optimized coating condition that allows a long-term maturation of human induced neurons in vitro.</p>}},
  author       = {{Kidnapillai, Srisaiyini and Ottosson, Daniella Rylander}},
  booktitle    = {{Neural Reprogramming: Methods and Protocols}},
  isbn         = {{978-1-0716-1601-7}},
  issn         = {{1940-6029}},
  keywords     = {{AAV; Action potential; Direct conversion; Fibroblast; GFP reporter; Glia; iNs; Intracerebral injections; iPSC; Lentivirus; Postsynaptic activity; Regeneration; Stem cells; Striatum; Transdifferentiation}},
  language     = {{eng}},
  pages        = {{183--199}},
  publisher    = {{Humana Press}},
  series       = {{Methods in Molecular Biology}},
  title        = {{Functional Assessment of Direct Reprogrammed Neurons In Vitro and In Vivo}},
  url          = {{http://dx.doi.org/10.1007/978-1-0716-1601-7_13}},
  doi          = {{10.1007/978-1-0716-1601-7_13}},
  volume       = {{2352}},
  year         = {{2021}},
}