Patterning of functional human astrocytes onto parylene-C/SiO2 substrates for the study of Ca2+ dynamics in astrocytic networks.
J Neural Eng
; 15(3): 036015, 2018 06.
Article
en En
| MEDLINE
| ID: mdl-29424361
ABSTRACT
OBJECTIVE:
Recent literature suggests that astrocytes form organized functional networks and communicate through transient changes in cytosolic Ca2+. Traditional techniques to investigate network activity, such as pharmacological blocking or genetic knockout, are difficult to restrict to individual cells. The objective of this work is to develop cell-patterning techniques to physically manipulate astrocytic interactions to enable the study of Ca2+ in astrocytic networks.APPROACH:
We investigate how an in vitro cell-patterning platform that utilizes geometric patterns of parylene-C on SiO2 can be used to physically isolate single astrocytes and small astrocytic networks. MAINRESULTS:
We report that single astrocytes are effectively isolated on 75 × 75 µm square parylene nodes, whereas multi-cellular astrocytic networks are isolated on larger nodes, with the mean number of astrocytes per cluster increasing as a function of node size. Additionally, we report that astrocytes in small multi-cellular clusters exhibit spatio-temporal clustering of Ca2+ transients. Finally, we report that the frequency and regularity of Ca2+ transients was positively correlated with astrocyte connectivity.SIGNIFICANCE:
The significance of this work is to demonstrate how patterning hNT astrocytes replicates spatio-temporal clustering of Ca2+ signalling that is observed in vivo but not in dissociated in vitro cultures. We therefore highlight the importance of the structure of astrocytic networks in determining ensemble Ca2+ behaviour.
Texto completo:
1
Banco de datos:
MEDLINE
Asunto principal:
Polímeros
/
Xilenos
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Diferenciación Celular
/
Astrocitos
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Dióxido de Silicio
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Señalización del Calcio
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Red Nerviosa
Límite:
Humans
Idioma:
En
Año:
2018
Tipo del documento:
Article