title: Human Cerebrospinal Fluid Promotes Neuronal Viability and Activity of Hippocampal Neuronal Circuits In Vitro.
abstract: For decades it has been hypothesized that molecules within the cerebrospinal fluid (CSF) diffuse into the brain parenchyma and influence the function of neurons. However, the functional consequences of CSF on neuronal circuits are largely unexplored and unknown. A major reason for this is the absence of appropriate neuronal in vitro model systems, and it is uncertain if neurons cultured in pure CSF survive and preserve electrophysiological functionality in vitro. In this article, we present an approach to address how human CSF (hCSF) influences neuronal circuits in vitro. We validate our approach by comparing the morphology, viability, and electrophysiological function of single neurons and at the network level in rat organotypic slice and primary neuronal cultures cultivated either in hCSF or in defined standard culture media. Our results demonstrate that rodent hippocampal slices and primary neurons cultured in hCSF maintain neuronal morphology and preserve synaptic transmission. Importantly, we show that hCSF increases neuronal viability and the number of electrophysiologically active neurons in comparison to the culture media. In summary, our data indicate that hCSF represents a physiological environment for neurons in vitro and a superior culture condition compared to the defined standard media. Moreover, this experimental approach paves the way to assess the functional consequences of CSF on neuronal circuits as well as suggesting a novel strategy for central nervous system (CNS) disease modeling.
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Marta
authors.initials: M
authors.lastname: Perez-Alcazar
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Georgia
authors.initials: G
authors.lastname: Culley
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Tim
authors.initials: T
authors.lastname: Lyckenvik
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Kristoffer
authors.initials: K
authors.lastname: Mobarrez
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Andreas
authors.initials: A
authors.lastname: Bjorefeldt
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Pontus
authors.initials: P
authors.lastname: Wasling
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Henrik
authors.initials: H
authors.lastname: Seth
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Frederik
authors.initials: F
authors.lastname: Asztely
authors.affiliation: Department of Neurology, Christian-Doppler-Klinik, Paracelsus Medical University Salzburg, Austria.
authors.forename: Andrea
authors.initials: A
authors.lastname: Harrer
authors.affiliation: Department of Geriatric Medicine, Christian-Doppler-Klinik, Paracelsus Medical University Salzburg, Austria.
authors.forename: Bernhard
authors.initials: B
authors.lastname: Iglseder
authors.affiliation: Institute of Molecular Regenerative Medicine, Paracelsus Medical UniversitySalzburg, Austria; Spinal Cord Injury and Tissue Regeneration Center Salzburg (SCI-TReCS), Paracelsus Medical UniversitySalzburg, Austria.
authors.forename: Ludwig
authors.initials: L
authors.lastname: Aigner
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of Gothenburg Gothenburg, Sweden.
authors.forename: Eric
authors.initials: E
authors.lastname: Hanse
authors.affiliation: Institute of Neuroscience and Physiology, The Sahlgrenska Academy, University of GothenburgGothenburg, Sweden; Institute of Molecular Regenerative Medicine, Paracelsus Medical UniversitySalzburg, Austria; Spinal Cord Injury and Tissue Regeneration Center Salzburg (SCI-TReCS), Paracelsus Medical UniversitySalzburg, Austria.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Illes
keywords: hippocampal neuronal cultures; hippocampal neuronal function; hippocampal neuronal survival; human cerebrospinal fluid; multi-electrode array technology; organotypic hippocampal slice cultures