Functional Assessment of Direct Reprogrammed Neurons In Vitro and In Vivo
(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.
(Less)
- author
- Kidnapillai, Srisaiyini LU and Ottosson, Daniella Rylander LU
- organization
- publishing date
- 2021
- 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
- 1064-3745
- 1940-6029
- ISBN
- 978-1-0716-1600-0
- 978-1-0716-1601-7
- 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
- 2024-08-11 03:14:15
@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 = {{978-1-0716-1600-0}}, issn = {{1064-3745}}, 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}}, }