Short-Term Free-Floating Slice Cultures from the Adult Human Brain
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Data
2019-11-01
Autores
Fernandes, Artur
Mendes, Niele Dias
Almeida, Glaucia Maria
Nogueira, Giovanna Orlovski
Machado, Carla de Moraes [UNESP]
Castro Horta-Junior, Jose de Anchieta de [UNESP]
Assirati Junior, Joao Alberto
Garcia-Cairasco, Norberto
Neder, Luciano
Sebollela, Adriano
Título da Revista
ISSN da Revista
Título de Volume
Editor
Journal Of Visualized Experiments
Resumo
Organotypic, or slice cultures, have been widely employed to model aspects of the central nervous system functioning in vitro. Despite the potential of slice cultures in neuroscience, studies using adult nervous tissue to prepare such cultures are still scarce, particularly those from human subjects. The use of adult human tissue to prepare slice cultures is particularly attractive to enhance the understanding of human neuropathologies, as they hold unique properties typical of the mature human brain lacking in slices produced from rodent (usually neonatal) nervous tissue. This protocol describes how to use brain tissue collected from living human donors submitted to resective brain surgery to prepare short-term, free-floating slice cultures. Procedures to maintain and perform biochemical and cell biology assays using these cultures are also presented. Representative results demonstrate that the typical human cortical lamination is preserved in slices after 4 days in vitro (DIV4), with expected presence of the main neural cell types. Moreover, slices at DIV4 undergo robust cell death when challenged with a toxic stimulus (H2O2), indicating the potential of this model to serve as a platform in cell death assays. This method, a simpler and cost-effective alternative to the widely used protocol using membrane inserts, is mainly recommended for running short-term assays aimed to unravel mechanisms of neurodegeneration behind age-associated brain diseases. Finally, although the protocol is devoted to using cortical tissue collected from patients submitted to surgical treatment of pharmacoresistant temporal lobe epilepsy, it is argued that tissue collected from other brain regions/conditions should also be considered as sources to produce similar free-floating slice cultures.
Descrição
Palavras-chave
Neuroscience, Issue 153, histoculture, neocortex, ex vivo, neurodegeneration, epilepsy, Alzheimer's
Como citar
Jove-journal Of Visualized Experiments. Cambridge: Journal Of Visualized Experiments, n. 153, 9 p., 2019.