{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,22]],"date-time":"2026-10-22T07:19:26Z","timestamp":1792653566604,"version":"4.5.0"},"reference-count":37,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2017,9,25]],"date-time":"2017-09-25T00:00:00Z","timestamp":1506297600000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2017,9,25]],"date-time":"2017-09-25T00:00:00Z","timestamp":1506297600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Pathophysiological investigation of CNS-related diseases, such as epilepsy or neurodegenerative disorders, largely relies on histological studies on human post mortem tissue, tissue obtained by biopsy or resective surgery and on studies using disease models including animal models, heterologous expression systems or cell culture based approaches. However, in general it remains elusive to what extent results obtained in model systems can be directly translated to the human brain, calling for strategies allowing validation or even primary investigation in live human CNS tissue. In the work reported here, we prepared human organotypic slice cultures from access tissue of resective epilepsy surgery. Employing different culture conditions, we systematically compared artificial culturing media versus human cerbrospinal fluid (hCSF) obtained from patients with normal pressure hydrocephalus (NPH). Presented data demonstrates sustained cortical neuronal survival including not only maintenance of typical cellular electrophysiological properties and activity, such as robust action potential generation and synaptic connectivity, but also preservation of tonic and phasic network activity up to several weeks<jats:italic>in vitro<\/jats:italic>. As clearly delineated by immunocytochemistry, single cell patch clamp and extracellular recordings, we find that in contrast to artificial culturing media, hCSF significantly enhances neuron viability and maintenance of network activity.<\/jats:p>","DOI":"10.1038\/s41598-017-12527-9","type":"journal-article","created":{"date-parts":[[2017,9,19]],"date-time":"2017-09-19T10:44:10Z","timestamp":1505817850000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":93,"title":["Human Cerebrospinal fluid promotes long-term neuronal viability and network function in human neocortical organotypic brain slice cultures"],"prefix":"10.1038","volume":"7","author":[{"given":"Niklas","family":"Schwarz","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ulrike B. S.","family":"Hedrich","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hannah","family":"Schwarz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harshad","family":"P.A.","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nele","family":"Dammeier","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Eva","family":"Auffenberg","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Francesco","family":"Bedogni","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"J\u00fcrgen B.","family":"Honegger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holger","family":"Lerche","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Thomas V.","family":"Wuttke","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henner","family":"Koch","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2017,9,25]]},"reference":[{"key":"12527_CR1","doi-asserted-by":"publisher","first-page":"547","DOI":"10.1038\/nature10648","volume":"480","author":"S Kriks","year":"2011","unstructured":"Kriks, S. et al. Dopamine neurons derived from human ES cells efficiently engraft in animal models of Parkinson\u2019s disease. Nature 480, 547\u2013551, https:\/\/doi.org\/10.1038\/nature10648 (2011).","journal-title":"Nature"},{"key":"12527_CR2","doi-asserted-by":"publisher","first-page":"785","DOI":"10.1016\/j.neuron.2013.05.029","volume":"78","author":"Y Zhang","year":"2013","unstructured":"Zhang, Y. et al. Rapid single-step induction of functional neurons from human pluripotent stem cells. Neuron 78, 785\u2013798, https:\/\/doi.org\/10.1016\/j.neuron.2013.05.029 (2013).","journal-title":"Neuron"},{"key":"12527_CR3","doi-asserted-by":"publisher","DOI":"10.1038\/nature12517","volume":"501","author":"MA Lancaster","year":"2013","unstructured":"Lancaster, M. A. et al. Cerebral organoids model human brain development and microcephaly. Nature 501, 373\u2212+, https:\/\/doi.org\/10.1038\/nature12517 (2013).","journal-title":"Nature"},{"key":"12527_CR4","doi-asserted-by":"publisher","first-page":"3","DOI":"10.1007\/s002210000395","volume":"133","author":"M D\u2019Esposito","year":"2000","unstructured":"D\u2019Esposito, M., Postle, B. R. & Rypma, B. Prefrontal cortical contributions to working memory: evidence from event-related fMRI studies. Experimental Brain Research 133, 3\u201311, https:\/\/doi.org\/10.1007\/s002210000395 (2000).","journal-title":"Experimental Brain Research"},{"key":"12527_CR5","doi-asserted-by":"publisher","first-page":"77","DOI":"10.1126\/science.280.5360.77","volume":"280","author":"RE Clark","year":"1998","unstructured":"Clark, R. E. & Squire, L. R. Classical conditioning and brain systems: the role of awareness. Science 280, 77\u201381 (1998).","journal-title":"Science"},{"key":"12527_CR6","doi-asserted-by":"publisher","unstructured":"Molnar, G. et al. Human pyramidal to interneuron synapses are mediated by multi-vesicular release and multiple docked vesicles. Elife 5, https:\/\/doi.org\/10.7554\/eLife.18167 (2016).","DOI":"10.7554\/eLife.18167"},{"key":"12527_CR7","doi-asserted-by":"publisher","first-page":"17197","DOI":"10.1523\/JNEUROSCI.3158-13.2013","volume":"33","author":"MB Verhoog","year":"2013","unstructured":"Verhoog, M. B. et al. Mechanisms underlying the rules for associative plasticity at adult human neocortical synapses. J Neurosci 33, 17197\u201317208, https:\/\/doi.org\/10.1523\/JNEUROSCI.3158-13.2013 (2013).","journal-title":"J Neurosci"},{"key":"12527_CR8","doi-asserted-by":"crossref","first-page":"7080","DOI":"10.1523\/JNEUROSCI.20-18-07080.2000","volume":"20","author":"H Beck","year":"2000","unstructured":"Beck, H., Goussakov, I. V., Lie, A., Helmstaedter, C. & Elger, C. E. Synaptic plasticity in the human dentate gyrus. J Neurosci 20, 7080\u20137086 (2000).","journal-title":"J Neurosci"},{"key":"12527_CR9","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pbio.1002007","volume":"12","author":"G Testa-Silva","year":"2014","unstructured":"Testa-Silva, G. et al. High bandwidth synaptic communication and frequency tracking in human neocortex. PLoS Biol 12, e1002007, https:\/\/doi.org\/10.1371\/journal.pbio.1002007 (2014).","journal-title":"PLoS Biol"},{"key":"12527_CR10","doi-asserted-by":"crossref","unstructured":"Sareen, D. et al. Targeting RNA Foci in iPSC-Derived Motor Neurons from ALS Patients with a C9ORF72 Repeat Expansion. Science Translational Medicine 5, doi:ARTN208ra14910.1126\/scitranslmed.3007529 (2013).","DOI":"10.1126\/scitranslmed.3007529"},{"key":"12527_CR11","doi-asserted-by":"publisher","first-page":"128","DOI":"10.1002\/ana.23897","volume":"74","author":"Y Liu","year":"2013","unstructured":"Liu, Y. et al. Dravet syndrome patient-derived neurons suggest a novel epilepsy mechanism. Ann Neurol 74, 128\u2013139, https:\/\/doi.org\/10.1002\/ana.23897 (2013).","journal-title":"Ann Neurol"},{"key":"12527_CR12","doi-asserted-by":"publisher","first-page":"86","DOI":"10.1016\/j.neuroscience.2015.07.086","volume":"305","author":"C Humpel","year":"2015","unstructured":"Humpel, C. Organotypic brain slice cultures: A review. Neuroscience 305, 86\u201398, https:\/\/doi.org\/10.1016\/j.neuroscience.2015.07.086 (2015).","journal-title":"Neuroscience"},{"key":"12527_CR13","doi-asserted-by":"publisher","first-page":"659","DOI":"10.1038\/nrd2617","volume":"7","author":"M Dragunow","year":"2008","unstructured":"Dragunow, M. The adult human brain in preclinical drug development. Nat Rev Drug Discov 7, 659\u2013666, https:\/\/doi.org\/10.1038\/nrd2617 (2008).","journal-title":"Nat Rev Drug Discov"},{"key":"12527_CR14","doi-asserted-by":"publisher","DOI":"10.1038\/srep24818","volume":"6","author":"M Andersson","year":"2016","unstructured":"Andersson, M. et al. Optogenetic control of human neurons in organotypic brain cultures. Sci Rep 6, 24818, https:\/\/doi.org\/10.1038\/srep24818 (2016).","journal-title":"Sci Rep"},{"key":"12527_CR15","doi-asserted-by":"publisher","first-page":"54","DOI":"10.1096\/fj.01-0504com","volume":"16","author":"RWH Verwer","year":"2002","unstructured":"Verwer, R. W. H. et al. Cells in human postmortem brain tissue slices remain alive for several weeks in culture. Faseb Journal 16, 54\u201360, https:\/\/doi.org\/10.1096\/fj.01-0504com (2002).","journal-title":"Faseb Journal"},{"key":"12527_CR16","doi-asserted-by":"publisher","first-page":"523","DOI":"10.1111\/bpa.12330","volume":"26","author":"RWH Verwer","year":"2016","unstructured":"Verwer, R. W. H. et al. Altered Loyalties of Neuronal Markers in Cultured Slices of Resected Human Brain Tissue. Brain Pathology 26, 523\u2013532, https:\/\/doi.org\/10.1111\/bpa.12330 (2016).","journal-title":"Brain Pathology"},{"key":"12527_CR17","doi-asserted-by":"publisher","first-page":"234","DOI":"10.1016\/j.jneumeth.2014.07.009","volume":"235","author":"E Eugene","year":"2014","unstructured":"Eugene, E. et al. An organotypic brain slice preparation from adult patients with temporal lobe epilepsy. J Neurosci Methods 235, 234\u2013244, https:\/\/doi.org\/10.1016\/j.jneumeth.2014.07.009 (2014).","journal-title":"J Neurosci Methods"},{"key":"12527_CR18","doi-asserted-by":"publisher","first-page":"429","DOI":"10.1016\/j.expneurol.2013.07.012","volume":"248","author":"N Daviaud","year":"2013","unstructured":"Daviaud, N., Garbayo, E., Schiller, P. C., Perez-Pinzon, M. & Montero-Menei, C. N. Organotypic cultures as tools for optimizing central nervous system cell therapies. Exp Neurol 248, 429\u2013440, https:\/\/doi.org\/10.1016\/j.expneurol.2013.07.012 (2013).","journal-title":"Exp Neurol"},{"key":"12527_CR19","doi-asserted-by":"publisher","first-page":"11678","DOI":"10.1523\/JNEUROSCI.4665-09.2010","volume":"30","author":"H Koch","year":"2010","unstructured":"Koch, H. et al. Prostaglandin E2-induced synaptic plasticity in neocortical networks of organotypic slice cultures. J Neurosci 30, 11678\u201311687, https:\/\/doi.org\/10.1523\/JNEUROSCI.4665-09.2010 (2010).","journal-title":"J Neurosci"},{"key":"12527_CR20","doi-asserted-by":"publisher","first-page":"11","DOI":"10.1007\/s00221-005-0069-z","volume":"168","author":"S Johansson","year":"2006","unstructured":"Johansson, S., Radesater, A. C., Cowburn, R. F., Thyberg, J. & Luthman, J. Modelling of amyloid beta-peptide induced lesions using roller-drum incubation of hippocampal slice cultures from neonatal rats. Exp Brain Res 168, 11\u201324, https:\/\/doi.org\/10.1007\/s00221-005-0069-z (2006).","journal-title":"Exp Brain Res"},{"key":"12527_CR21","doi-asserted-by":"publisher","first-page":"5084","DOI":"10.1523\/JNEUROSCI.0258-16.2016","volume":"36","author":"R Novotny","year":"2016","unstructured":"Novotny, R. et al. Conversion of Synthetic Abeta to In Vivo Active Seeds and Amyloid Plaque Formation in a Hippocampal Slice Culture Model. J Neurosci 36, 5084\u20135093, https:\/\/doi.org\/10.1523\/JNEUROSCI.0258-16.2016 (2016).","journal-title":"J Neurosci"},{"key":"12527_CR22","doi-asserted-by":"publisher","first-page":"87","DOI":"10.1002\/cne.903070109","volume":"307","author":"M Caeser","year":"1991","unstructured":"Caeser, M. & Aertsen, A. Morphological organization of rat hippocampal slice cultures. J Comp Neurol 307, 87\u2013106, https:\/\/doi.org\/10.1002\/cne.903070109 (1991).","journal-title":"J Comp Neurol"},{"key":"12527_CR23","doi-asserted-by":"publisher","first-page":"85","DOI":"10.1046\/j.1460-9568.2003.02728.x","volume":"18","author":"KV Moser","year":"2003","unstructured":"Moser, K. V., Schmidt-Kastner, R., Hinterhuber, H. & Humpel, C. Brain capillaries and cholinergic neurons persist in organotypic brain slices in the absence of blood flow. Eur J Neurosci 18, 85\u201394 (2003).","journal-title":"Eur J Neurosci"},{"key":"12527_CR24","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1113\/jphysiol.2003.039099","volume":"550","author":"A De Simoni","year":"2003","unstructured":"De Simoni, A., Griesinger, C. B. & Edwards, F. A. Development of rat CA1 neurones in acute versus organotypic slices: role of experience in synaptic morphology and activity. J Physiol 550, 135\u2013147, https:\/\/doi.org\/10.1113\/jphysiol.2003.039099 (2003).","journal-title":"J Physiol"},{"key":"12527_CR25","doi-asserted-by":"publisher","DOI":"10.15252\/msb.20145508","volume":"10","author":"V Busskamp","year":"2014","unstructured":"Busskamp, V. et al. Rapid neurogenesis through transcriptional activation in human stem cells. Molecular systems biology 10, 760, https:\/\/doi.org\/10.15252\/msb.20145508 (2014).","journal-title":"Molecular systems biology"},{"key":"12527_CR26","doi-asserted-by":"publisher","first-page":"1836","DOI":"10.1038\/nprot.2012.116","volume":"7","author":"Y Shi","year":"2012","unstructured":"Shi, Y., Kirwan, P. & Livesey, F. J. Directed differentiation of human pluripotent stem cells to cerebral cortex neurons and neural networks. Nat Protoc 7, 1836\u20131846, https:\/\/doi.org\/10.1038\/nprot.2012.116 (2012).","journal-title":"Nat Protoc"},{"key":"12527_CR27","doi-asserted-by":"publisher","unstructured":"Bardy, C. et al. Neuronal medium that supports basic synaptic functions and activity of human neurons in vitro. Proceedings of the National Academy of Sciences of the United States of America, https:\/\/doi.org\/10.1073\/pnas.1504393112 (2015).","DOI":"10.1073\/pnas.1504393112"},{"key":"12527_CR28","doi-asserted-by":"publisher","DOI":"10.1186\/1743-8454-7-1","volume":"7","author":"JG Veening","year":"2010","unstructured":"Veening, J. G. & Barendregt, H. P. The regulation of brain states by neuroactive substances distributed via the cerebrospinal fluid; a review. Cerebrospinal Fluid Res 7, 1, https:\/\/doi.org\/10.1186\/1743-8454-7-1 (2010).","journal-title":"Cerebrospinal Fluid Res"},{"key":"12527_CR29","doi-asserted-by":"publisher","DOI":"10.3389\/fncel.2016.00054","volume":"10","author":"M Perez-Alcazar","year":"2016","unstructured":"Perez-Alcazar, M. et al. Human Cerebrospinal Fluid Promotes Neuronal Viability and Activity of Hippocampal Neuronal Circuits In Vitro. Front Cell Neurosci 10, 54, https:\/\/doi.org\/10.3389\/fncel.2016.00054 (2016).","journal-title":"Front Cell Neurosci"},{"key":"12527_CR30","doi-asserted-by":"publisher","first-page":"15","DOI":"10.1111\/j.1749-6632.1988.tb31808.x","volume":"531","author":"RM Post","year":"1988","unstructured":"Post, R. M., Rubinow, D. R., Kling, M. A., Berrettini, W. & Gold, P. W. Neuroactive substances in cerebrospinal fluid. Normal and pathological regulatory mechanisms. Ann N Y Acad Sci 531, 15\u201328 (1988).","journal-title":"Ann N Y Acad Sci"},{"key":"12527_CR31","doi-asserted-by":"publisher","first-page":"1335","DOI":"10.1093\/cercor\/bhm165","volume":"18","author":"N Heck","year":"2008","unstructured":"Heck, N. et al. Activity-dependent regulation of neuronal apoptosis in neonatal mouse cerebral cortex. Cereb Cortex 18, 1335\u20131349, https:\/\/doi.org\/10.1093\/cercor\/bhm165 (2008).","journal-title":"Cereb Cortex"},{"key":"12527_CR32","doi-asserted-by":"publisher","first-page":"231","DOI":"10.1113\/jphysiol.2014.284711","volume":"593","author":"A Bjorefeldt","year":"2015","unstructured":"Bjorefeldt, A. et al. Human cerebrospinal fluid increases the excitability of pyramidal neurons in the in vitro brain slice. J Physiol 593, 231\u2013243, https:\/\/doi.org\/10.1113\/jphysiol.2014.284711 (2015).","journal-title":"J Physiol"},{"key":"12527_CR33","doi-asserted-by":"crossref","first-page":"414","DOI":"10.55782\/ane-2008-1708","volume":"68","author":"J Skipor","year":"2008","unstructured":"Skipor, J. & Thiery, J. C. The choroid plexus\u2013cerebrospinal fluid system: undervaluated pathway of neuroendocrine signaling into the brain. Acta Neurobiol Exp (Wars) 68, 414\u2013428 (2008).","journal-title":"Acta Neurobiol Exp (Wars)"},{"key":"12527_CR34","doi-asserted-by":"publisher","first-page":"84","DOI":"10.1210\/endo.143.1.8585","volume":"143","author":"H Tricoire","year":"2002","unstructured":"Tricoire, H., Locatelli, A., Chemineau, P. & Malpaux, B. Melatonin enters the cerebrospinal fluid through the pineal recess. Endocrinology 143, 84\u201390, https:\/\/doi.org\/10.1210\/en.143.1.84 (2002).","journal-title":"Endocrinology"},{"key":"12527_CR35","doi-asserted-by":"publisher","first-page":"546","DOI":"10.1002\/ana.21808","volume":"66","author":"F Otto","year":"2009","unstructured":"Otto, F. et al. Cerebrospinal fluid of brain trauma patients inhibits in vitro neuronal network function via NMDA receptors. Ann Neurol 66, 546\u2013555, https:\/\/doi.org\/10.1002\/ana.21808 (2009).","journal-title":"Ann Neurol"},{"key":"12527_CR36","doi-asserted-by":"publisher","unstructured":"Bagheri, M., Rezakhani, A., Roghani, M., Joghataei, M. T. & Mohseni, S. Protocol for Three-dimensional Confocal Morphometric Analysis of Astrocytes. J Vis Exp, e53113, https:\/\/doi.org\/10.3791\/53113 (2015).","DOI":"10.3791\/53113"},{"key":"12527_CR37","doi-asserted-by":"publisher","first-page":"1077","DOI":"10.1152\/physrev.00041.2013","volume":"94","author":"M Pekny","year":"2014","unstructured":"Pekny, M. & Pekna, M. Astrocyte reactivity and reactive astrogliosis: costs and benefits. Physiol Rev 94, 1077\u20131098, https:\/\/doi.org\/10.1152\/physrev.00041.2013 (2014).","journal-title":"Physiol Rev"}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-12527-9","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-12527-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-12527-9.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,6,27]],"date-time":"2024-06-27T01:51:32Z","timestamp":1719453092000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-017-12527-9"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,9,25]]},"references-count":37,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2017,12]]}},"alternative-id":["12527"],"URL":"https:\/\/doi.org\/10.1038\/s41598-017-12527-9","relation":{},"ISSN":["2045-2322"],"issn-type":[{"value":"2045-2322","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,9,25]]},"assertion":[{"value":"5 April 2017","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 September 2017","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 September 2017","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare that they have no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing Interests"}}],"article-number":"12249"}}