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Cano-Abad 2011 Int J Dev Neurosci

From Bioblast
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Cano-Abad MF1, Herrera-Peco I, Sola RG, Pastor J, García-Navarrete E, Moro RC, Pizzo P, Ruiz-Nuño A (2011) Morphological and functional characterization of human cells from tissue coming from epileptic patients under surgery. Int J Dev Neurosci 29:121-9.

» PMID: 21238565

Cano-Abad MF, Herrera-Peco I, Sola RG, Pastor J, Garcia-Navarrete E, Moro RC, Pizzo P, Ruiz-Nuno A (2011) Int J Dev Neurosci

Abstract: Primary brain cell cultures are a useful tool for understanding the physiopathology of epilepsy and for searching new potential antiepileptic drugs. These cell types are usually prepared from murine species and few human models have been described. The main goal of this study is the establishment of experimental conditions to isolate and culture neurons and astrocytes from human brain and to test its functionality. The tissues came from antiepileptic drug-resistant epileptic patients undergoing surgery. Human neurons and astrocytes were isolated following an enzymatic and mechanical dissociation protocol. Cultures were viable for 3-6 weeks. Cytological characterization was performed by immunocytochemistry using specific antibodies against both neuron (anti-NeuN) and astrocyte (anti-GFAP) protein markers. In order to test their viability and functionality, cells were loaded with the fluorescent calcium probe fura-2 and variations in cytosolic calcium concentrations ([Ca2+]c) were measured by cell imaging. [Ca2+]c increases were evoked upon cell stimulation with high Ka+ (KCl 75 mM), glutamate (500 μM) or bicuculline (100 μM). Interestingly, spontaneous [Ca2+]c transients were also observed in some neuron-like cells. A novel unreported finding in this study has been the incorporation of human serum that was critical for cell functionality. The setting of these human cultures open the opportunity to new insights on culture and calcium signalling studies on the mechanism(s) of cell resistance to antiepileptic drugs, as well as to studies on plasticity, maturation and possible neurite emission for graft studies.

Copyright © 2011 ISDN. Published by Elsevier Ltd. All rights reserved.


Labels: Pathology: Other 

Organism: Human  Tissue;cell: Nervous system