Description Of A Six-Step Approach To Gain Higher Quality Results From Organotypic Hippocampal Brain Slices in A Traumatic Brain Injury Model

Introduction: Organotypic Hippocampal Brain Slices (OHBS) provide an advantageous alternative to in vivo models to scrutinize Traumatic Brain Injury (TBI). We followed a well-established TBI protocol, but noticed that several factors may influence the results in such a setup. Here, we describe a structured approach to generate more comparable results and discuss why specific eligibility criteria should be applied. Methods: We defined necessary checkpoints and developed inclusion and exclusion criteria that take the observed variation in such a model into consideration. Objective measures include the identification and exclusion of pre-damaged slices and outliers. Six steps were outlined in this study. Results: A six-step approach to enhance comparability is proposed and summarized in a flowchart. We applied the suggested measures to data derived from our TBI-experiments examining the impact of three different interventions in 1459 OHBS. Our exemplary results show that through equal requirements set for all slices more precise findings are ensured. Conclusion: Results in a TBI experiment on OHBS should be analyzed critically as inhomogeneities may occur. In order to ensure more precise findings, a structured approach of comparing the results should be followed. Further research is recommended to confirm and further develop this framework.

[1]  R. Rossaint,et al.  Argon attenuates the emergence of secondary injury after traumatic brain injury within a 2-hour incubation period compared to desflurane: an in vitro study , 2017, Medical gas research.

[2]  B. Bahr,et al.  Blast waves from detonated military explosive reduce GluR1 and synaptophysin levels in hippocampal slice cultures , 2016, Experimental Neurology.

[3]  Jian Wang,et al.  Organotypic Hippocampal Slices as Models for Stroke and Traumatic Brain Injury , 2016, Molecular Neurobiology.

[4]  Tapan P. Patel,et al.  Isolated Primary Blast Inhibits Long-Term Potentiation in Organotypic Hippocampal Slice Cultures. , 2016, Journal of neurotrauma.

[5]  R. Rossaint,et al.  Desflurane impairs outcome of organotypic hippocampal slices in an in vitro model of traumatic brain injury , 2016, Medical gas research.

[6]  C. Humpel,et al.  ORGANOTYPIC BRAIN SLICE CULTURES: A REVIEW , 2015, Neuroscience.

[7]  S. Kurpad,et al.  Effects of Blast Overpressure on Neurons and Glial Cells in Rat Organotypic Hippocampal Slice Cultures , 2015, Front. Neurol..

[8]  D. Shreiber,et al.  Neuroprotection by genipin against reactive oxygen and reactive nitrogen species-mediated injury in organotypic hippocampal slice cultures , 2014, Brain Research.

[9]  R. Dickinson,et al.  Neuroprotection against Traumatic Brain Injury by Xenon, but Not Argon, Is Mediated by Inhibition at the N-Methyl-D-Aspartate Receptor Glycine Site , 2013, Anesthesiology.

[10]  Mohammad Wahid Ansari,et al.  The legal status of in vitro embryos , 2014 .

[11]  R. Rossaint,et al.  Argon reduces neurohistopathological damage and preserves functional recovery after cardiac arrest in rats. , 2013, British journal of anaesthesia.

[12]  D. Hovda,et al.  Potentially neuroprotective gene modulation in an in vitro model of mild traumatic brain injury , 2013, Molecular and Cellular Biochemistry.

[13]  M. Gillette,et al.  Over a Century of Neuron Culture: From the Hanging Drop to Microfluidic Devices , 2012, The Yale journal of biology and medicine.

[14]  M. Vizcaychipi,et al.  The protective profile of argon, helium, and xenon in a model of neonatal asphyxia in rats* , 2012, Critical care medicine.

[15]  R. Rossaint,et al.  Dexmedetomidine is neuroprotective in an in vitro model for traumatic brain injury , 2012, BMC Neurology.

[16]  S. Weber,et al.  A simple method for measuring organotypic tissue slice culture thickness , 2011, Journal of Neuroscience Methods.

[17]  R. Rossaint,et al.  Neuroprotective properties of levosimendan in an in vitro model of traumatic brain injury , 2010, BMC neurology.

[18]  Antonio Belli,et al.  Transcriptomics of traumatic brain injury: gene expression and molecular pathways of different grades of insult in a rat organotypic hippocampal culture model. , 2010, Journal of neurotrauma.

[19]  R. Rossaint,et al.  Argon: Neuroprotection in in vitro models of cerebral ischemia and traumatic brain injury , 2009, Critical care.

[20]  R. Rossaint,et al.  Propofol: neuroprotection in an in vitro model of traumatic brain injury , 2009, Critical care.

[21]  M. Maze,et al.  The neuroprotective effects of xenon and helium in an in vitro model of traumatic brain injury* , 2008, Critical care medicine.

[22]  Mark R Bowlby,et al.  Brain slices as models for neurodegenerative disease and screening platforms to identify novel therapeutics. , 2007, Current neuropharmacology.

[23]  L. M. Yu,et al.  Preparation of organotypic hippocampal slice cultures: interface method , 2006, Nature Protocols.

[24]  P. Caroni,et al.  Preparation of organotypic hippocampal slice cultures for long-term live imaging , 2006, Nature Protocols.

[25]  L. Sundstrom,et al.  An in vitro model of traumatic brain injury utilising two-dimensional stretch of organotypic hippocampal slice cultures , 2006, Journal of Neuroscience Methods.

[26]  A. Luft,et al.  Stages of motor skill learning , 2005, Molecular Neurobiology.

[27]  F. Poulsen,et al.  Organotypic hippocampal slice cultures for studies of brain damage, neuroprotection and neurorepair. , 2005, Current drug targets. CNS and neurological disorders.

[28]  A. Wood,et al.  Functional validation of adult hippocampal organotypic cultures as an in vitro model of brain injury , 2004, Brain Research.

[29]  L. Sundstrom,et al.  A tissue level tolerance criterion for living brain developed with an in vitro model of traumatic mechanical loading. , 2003, Stapp car crash journal.

[30]  P. Carlen,et al.  Methods to induce primary and secondary traumatic damage in organotypic hippocampal slice cultures. , 2000, Brain research. Brain research protocols.

[31]  D F Meaney,et al.  In vitro central nervous system models of mechanically induced trauma: a review. , 1998, Journal of neurotrauma.

[32]  D. Debanne,et al.  Organotypic slice cultures: a technique has come of age , 1997, Trends in Neurosciences.

[33]  D. Muller,et al.  A simple method for organotypic cultures of nervous tissue , 1991, Journal of Neuroscience Methods.

[34]  J. D. Macklis,et al.  Progressive incorporation of propidium iodide in cultured mouse neurons correlates with declining electrophysiological status: a fluorescence scale of membrane integrity , 1990, Journal of Neuroscience Methods.

[35]  B. Gähwiler.  Organotypic monolayer cultures of nervous tissue , 1981, Journal of Neuroscience Methods.

[36]  Hogue Mj.  Review of studies of human fetal brain cells in tissue cultures. , 1952 .

[37]  R. G. Harrison.  The outgrowth of the nerve fiber as a mode of protoplasmic movement. , 1910, The Journal of experimental zoology.

[38]  M. J. Greenman,et al.  Observations of the living developing nerve fiber , 1907 .

[39]  S. Thompson,et al.  Preparation of Organotypic Hippocampal Slice Cultures , 2006 .

[40]  B. Bahr,et al.  Stable maintenance of glutamate receptors and other synaptic components in long‐term hippocampal slices , 1995, Hippocampus.

[41]  M. J. Hogue.  Review of studies of human fetal brain cells in tissue cultures. , 1952, Etudes neo-natales. Neo-natal studies.