Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays

Given the critical roles of astrocytes in neuroinflammation and neurological diseases, models for studying human astrocyte biology are in increasing demand. Here, we present a protocol to isolate human astrocytes from induced pluripotent stem cell (iPSC)-based cultures, neural organoids, and primary...

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Autores principales: Barbar, Lilianne, Rusielewicz, Tomasz, Zimmer, Matthew, Kalpana, Kriti, Fossati, Valentina
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2020
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757411/
https://www.ncbi.nlm.nih.gov/pubmed/33377066
http://dx.doi.org/10.1016/j.xpro.2020.100172
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author Barbar, Lilianne
Rusielewicz, Tomasz
Zimmer, Matthew
Kalpana, Kriti
Fossati, Valentina
author_facet Barbar, Lilianne
Rusielewicz, Tomasz
Zimmer, Matthew
Kalpana, Kriti
Fossati, Valentina
author_sort Barbar, Lilianne
collection PubMed
description Given the critical roles of astrocytes in neuroinflammation and neurological diseases, models for studying human astrocyte biology are in increasing demand. Here, we present a protocol to isolate human astrocytes from induced pluripotent stem cell (iPSC)-based cultures, neural organoids, and primary tissue, using the surface marker CD49f. Moreover, we provide protocols for in vitro co-cultures of human iPSC-derived neurons and astrocytes, as well as for neurotoxicity assays that expose neurons to conditioned media from reactive astrocytes. For complete details on the use and execution of this protocol, please refer to Barbar et al. (2020).
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spelling pubmed-77574112020-12-28 Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays Barbar, Lilianne Rusielewicz, Tomasz Zimmer, Matthew Kalpana, Kriti Fossati, Valentina STAR Protoc Protocol Given the critical roles of astrocytes in neuroinflammation and neurological diseases, models for studying human astrocyte biology are in increasing demand. Here, we present a protocol to isolate human astrocytes from induced pluripotent stem cell (iPSC)-based cultures, neural organoids, and primary tissue, using the surface marker CD49f. Moreover, we provide protocols for in vitro co-cultures of human iPSC-derived neurons and astrocytes, as well as for neurotoxicity assays that expose neurons to conditioned media from reactive astrocytes. For complete details on the use and execution of this protocol, please refer to Barbar et al. (2020). Elsevier 2020-11-10 /pmc/articles/PMC7757411/ /pubmed/33377066 http://dx.doi.org/10.1016/j.xpro.2020.100172 Text en © 2020 The Author(s) http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Barbar, Lilianne
Rusielewicz, Tomasz
Zimmer, Matthew
Kalpana, Kriti
Fossati, Valentina
Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays
title Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays
title_full Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays
title_fullStr Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays
title_full_unstemmed Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays
title_short Isolation of Human CD49f(+) Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays
title_sort isolation of human cd49f(+) astrocytes and in vitro ipsc-based neurotoxicity assays
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7757411/
https://www.ncbi.nlm.nih.gov/pubmed/33377066
http://dx.doi.org/10.1016/j.xpro.2020.100172
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