Free-floating adult human brain-derived slice cultures as a model to study the neuronal impact of Alzheimer’s disease-associated Aβ oligomers

Publication date: Available online 31 May 2018
Source:Journal of Neuroscience Methods
Author(s): Niele D. Mendes, Artur Fernandes, Glaucia M. Almeida, Luis E. Santos, Maria Clara Selles, Natalia Lyra-Silva, Carla M. Machado, José A.C. Horta-Júnior, Paulo R. Louzada, Fernanda G. De Felice, Soniza Alvez-Leon, Jorge Marcondes, João Alberto Assirati, Caio M. Matias, William L. Klein, Norberto Garcia-Cairasco, Sergio T. Ferreira, Luciano Neder, Adriano Sebollela
BackgroundSlice cultures have been prepared from several organs. With respect to the brain, advantages of slice cultures over dissociated cell cultures include maintenance of the cytoarchitecture and neuronal connectivity. Slice cultures from adult human brain have been reported and constitute a promising method to study neurological diseases. Despite this potential, few studies have characterized in detail cell survival and function along time in short-term, free-floating cultures.New MethodWe used tissue from adult human brain cortex from patients undergoing temporal lobectomy to prepare 200 μm-thick slices. Along the period in culture, we evaluated neuronal survival, histological modifications, and neurotransmitter release. The toxicity of Alzheimer's-associated Aβ oligomers (AβOs) to cultured slices was also analyzed.ResultsNeurons in human brain slices remain viable and neurochemically active for at least four days in vitro, which allowed detection of binding of AβOs. We further found that slices exposed to AβOs presented elevated levels of hyperphosphorylated Tau, a hallmark of Alzheimer's disease.Comparison with Existing Method(s)Although slice cultures from adult human brain have been previously prepared, this is the first report to analyze cell viability and neuronal activity in short-term free-floating cultures as a function of days in vitro.ConclusionsOnce surgical tissue is available, the current protocol is easy to perform and produces functional slices from adult human brain. These slice cultures may represent a preferred model for translational studies of neurodegenerative disorders when long term culturing in not required, as in investigations on AβO neurotoxicity.



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