Abstract
Cell cultures are characterized by their simplicity, controllability, and ability to provide detailed basic information on how a particular cell population responds to specific stimuli or insult. These characteristics led to their extensive application in the study of molecular interactions and represent a valuable tool in the study of different pathologies. However, due to the lack of interactions between the different components that form an in vivo system, the results obtained in pure cell cultures not always translate what occurs in vivo. In this context, the use of co-cultures has the advantage of allowing the study of interactions between different types of cells present in a tissue, which in many situations are determinant for the effects obtained. The present study aimed to characterize cortical neuron–glia and neuron-enriched primary cultures and evaluate their response to an ischemic insult. Cell viability was assessed by the MTT assay and cell number/phenotype was analyzed by immunocytochemistry in control cultures and in cells subjected to 4 h of oxygen and glucose deprivation. The results obtained demonstrate that astrocytes have a substantial impact on the injury induced by an ischemic insult, thus suggesting that the crosstalk between glia and neurons is crucial to the neuronal protection in conditions of ischemia.
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Abbreviations
- BBB:
-
Blood–brain barrier
- CNS:
-
Central nervous system
- FBS:
-
Fetal bovine serum
- GFAP:
-
Glial fibrillary acidic protein
- HBSS:
-
Hank’s buffered salt solution
- Iba1:
-
Ionized calcium-binding adaptor molecule 1
- IS:
-
Ischemic stroke
- MAP2:
-
Microtubule-associated protein 2
- NBM:
-
Neurobasal medium
- MTT:
-
3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
- OGD:
-
Oxygen and glucose deprivation
- PBS:
-
Phosphate buffered saline
- PBS-T:
-
Phosphate buffered saline with 0.1% Tween
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Acknowledgements
The authors would like to thank Dr. Ana Saavedra (Facultat de Medicina, Institut de Neurociències, Universitat de Barcelona) for her careful and critical reading of the manuscript and also acknowledge the funding support provided by FEDER funds through the POCI - COMPETE 2020 - Operational Programme Competitiveness and Internationalisation in Axis I - Strengthening research, technological development and innovation (Project POCI-01-0145-FEDER-007491), and National Funds by FCT - Foundation for Science and Technology (Project UID/Multi/00709/2013).
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Roque, C., Baltazar, G. Impact of Astrocytes on the Injury Induced by In Vitro Ischemia. Cell Mol Neurobiol 37, 1521–1528 (2017). https://doi.org/10.1007/s10571-017-0483-3
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DOI: https://doi.org/10.1007/s10571-017-0483-3