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Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells

Astrocytes play an important role in cell–cell signaling in the mammalian central nervous system. The ability of astrocytes to communicate with surrounding cells through gap-junctional coupling or signaling via the release of transmitters makes characterization of these cells difficult in vitro and...

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Detalles Bibliográficos
Autores principales: Lee, William, Malarkey, Erik B., Reyes, Reno C., Parpura, Vladimir
Formato: Texto
Lenguaje:English
Publicado: Frontiers Research Foundation 2008
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2610544/
https://www.ncbi.nlm.nih.gov/pubmed/19129909
http://dx.doi.org/10.3389/neuro.16.002.2008
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author Lee, William
Malarkey, Erik B.
Reyes, Reno C.
Parpura, Vladimir
author_facet Lee, William
Malarkey, Erik B.
Reyes, Reno C.
Parpura, Vladimir
author_sort Lee, William
collection PubMed
description Astrocytes play an important role in cell–cell signaling in the mammalian central nervous system. The ability of astrocytes to communicate with surrounding cells through gap-junctional coupling or signaling via the release of transmitters makes characterization of these cells difficult in vitro and even more so in vivo. To simplify the complexity of common in vitro systems, introduced by intercellular communication between astrocytes, we developed a novel cell culturing method, in which purified rat visual cortical astrocytes were grown in spatially defined cell-adhesion wells which we termed micropits. We showed that astrocytes cultured in micropit regions were viable and exhibited similar characteristics of Ca(2+) dynamics and astrocytic marker expression to those of cells cultured in non-micropit regions. Examination of intracellular Ca(2+) oscillations in solitary astrocytes cultured in micropits revealed less variable oscillations than those of non-micropit grouped astrocytes, which were in contact with their neighbors. Solitary cells in micropit regions can undergo ATP-mediated astrocyte-microglia signaling, demonstrating that this culturing method can also be used to investigate glial–glial interactions in a spatially well-defined microenvironment.
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spelling pubmed-26105442009-01-07 Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells Lee, William Malarkey, Erik B. Reyes, Reno C. Parpura, Vladimir Front Neuroengineering Neuroscience Astrocytes play an important role in cell–cell signaling in the mammalian central nervous system. The ability of astrocytes to communicate with surrounding cells through gap-junctional coupling or signaling via the release of transmitters makes characterization of these cells difficult in vitro and even more so in vivo. To simplify the complexity of common in vitro systems, introduced by intercellular communication between astrocytes, we developed a novel cell culturing method, in which purified rat visual cortical astrocytes were grown in spatially defined cell-adhesion wells which we termed micropits. We showed that astrocytes cultured in micropit regions were viable and exhibited similar characteristics of Ca(2+) dynamics and astrocytic marker expression to those of cells cultured in non-micropit regions. Examination of intracellular Ca(2+) oscillations in solitary astrocytes cultured in micropits revealed less variable oscillations than those of non-micropit grouped astrocytes, which were in contact with their neighbors. Solitary cells in micropit regions can undergo ATP-mediated astrocyte-microglia signaling, demonstrating that this culturing method can also be used to investigate glial–glial interactions in a spatially well-defined microenvironment. Frontiers Research Foundation 2008-12-12 /pmc/articles/PMC2610544/ /pubmed/19129909 http://dx.doi.org/10.3389/neuro.16.002.2008 Text en Copyright: © 2008 Lee, Malarkey, Reyes and Parpura. http://www.frontiersin.org/licenseagreement This is an open-access article subject to an exclusive license agreement between the authors and the Frontiers Research Foundation, which permits unrestricted use, distribution, and reproduction in any medium, provided the original authors and source are credited.
spellingShingle Neuroscience
Lee, William
Malarkey, Erik B.
Reyes, Reno C.
Parpura, Vladimir
Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells
title Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells
title_full Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells
title_fullStr Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells
title_full_unstemmed Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells
title_short Micropit: A New Cell Culturing Approach for Characterization of Solitary Astrocytes and Small Networks of these Glial Cells
title_sort micropit: a new cell culturing approach for characterization of solitary astrocytes and small networks of these glial cells
topic Neuroscience
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2610544/
https://www.ncbi.nlm.nih.gov/pubmed/19129909
http://dx.doi.org/10.3389/neuro.16.002.2008
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