{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,29]],"date-time":"2025-10-29T06:24:57Z","timestamp":1761719097803,"version":"build-2065373602"},"reference-count":49,"publisher":"MDPI AG","issue":"1","license":[{"start":{"date-parts":[[2022,1,4]],"date-time":"2022-01-04T00:00:00Z","timestamp":1641254400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Ministry of Science and Culture of Lower Saxony","award":["Niedersachsen\u2010Research Network on Neuroinfectiology (N\u2010RENNT)"],"award-info":[{"award-number":["Niedersachsen\u2010Research Network on Neuroinfectiology (N\u2010RENNT)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>The host\u2013pathogen interaction during meningitis can be investigated with blood-cerebrospinal-fluid-barrier (BCSFB) cell culture models. They are commonly handled under atmospheric oxygen conditions (19\u201321% O2), although the physiological oxygen conditions are significantly lower in cerebrospinal fluid (CSF) (7\u20138% O2). We aimed to characterize oxygen levels in a Streptococcus (S.) suis-infected BCSFB model with transmigrating neutrophils. A BCSFB model with human choroid plexus epithelial cells growing on transwell-filters was used. The upper \u201cblood\u201d-compartment was infected and blood-derived neutrophils were added. S. suis and neutrophils transmigrated through the BCSFB into the \u201cCSF\u201d-compartment. Here, oxygen and pH values were determined with the non-invasive SensorDish\u00ae reader. Slight orbital shaking improved the luminescence-based measurement technique for detecting free oxygen. In the non-infected BCSFB model, an oxygen value of 7% O2 was determined. However, with S. suis and transmigrating neutrophils, the oxygen value significantly decreased to 2% O2. The pH level decreased slightly in all groups. In conclusion, we characterized oxygen levels in the BCSFB model and demonstrated the oxygen consumption by cells and bacteria. Oxygen values in the non-infected BCSFB model are comparable to in vivo values determined in pigs in the CSF. Infection and transmigrating neutrophils decrease the oxygen value to lower values.<\/jats:p>","DOI":"10.3390\/cells11010151","type":"journal-article","created":{"date-parts":[[2022,1,5]],"date-time":"2022-01-05T20:42:16Z","timestamp":1641415336000},"page":"151","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Characterization of Oxygen Levels in an Uninfected and Infected Human Blood-Cerebrospinal-Fluid-Barrier Model"],"prefix":"10.3390","volume":"11","author":[{"given":"Alexander","family":"Martens","sequence":"first","affiliation":[{"name":"Department of Biochemistry, University of Veterinary Medicine Hannover, 30559 Hannover, Germany"},{"name":"Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4516-7838","authenticated-orcid":false,"given":"Nicole","family":"de Buhr","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Veterinary Medicine Hannover, 30559 Hannover, Germany"},{"name":"Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7312-0682","authenticated-orcid":false,"given":"Hiroshi","family":"Ishikawa","sequence":"additional","affiliation":[{"name":"Laboratory of Clinical Regenerative Medicine, Department of Neurosurgery, University of Tsukuba, Tsukuba-City, Inaraki 305-8575, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Horst","family":"Schroten","sequence":"additional","affiliation":[{"name":"Department of Pediatrics, Pediatric Infectious Diseases, Medical Faculty Mannheim, Heidelberg University, 68167 Mannheim, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7712-9767","authenticated-orcid":false,"given":"Maren","family":"von K\u00f6ckritz-Blickwede","sequence":"additional","affiliation":[{"name":"Department of Biochemistry, University of Veterinary Medicine Hannover, 30559 Hannover, Germany"},{"name":"Research Center for Emerging Infections and Zoonoses (RIZ), University of Veterinary Medicine Hannover, 30559 Hannover, Germany"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2022,1,4]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"697","DOI":"10.1016\/j.arcmed.2005.06.009","article-title":"Resistance to Antibiotics: Are We in the Post-Antibiotic Era?","volume":"36","author":"Alanis","year":"2005","journal-title":"Arch. Med. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1016\/S1473-3099(18)30605-4","article-title":"Attributable deaths and disability-adjusted life-years caused by infections with antibiotic-resistant bacteria in the EU and the European Economic Area in 2015: A population-level modelling analysis","volume":"19","author":"Cassini","year":"2019","journal-title":"Lancet Infect. Dis."},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"B\u00e9dard, P., Gauvin, S., Ferland, K., Caneparo, C., Pellerin, \u00c8., Chabaud, S., and Bolduc, S. (2020). Innovative Human Three-Dimensional Tissue-Engineered Models as an Alternative to Animal Testing. Bioengineering, 7.","DOI":"10.3390\/bioengineering7030115"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6","DOI":"10.3389\/fphar.2018.00006","article-title":"Three-Dimensional in Vitro Cell Culture Models in Drug Discovery and Drug Repositioning","volume":"9","author":"Langhans","year":"2018","journal-title":"Front. Pharmacol."},{"key":"ref_5","doi-asserted-by":"crossref","unstructured":"Zi\u00f3\u0142kowska-Suchanek, I. (2021). Mimicking Tumor Hypoxia in Non-Small Cell Lung Cancer Employing Three-Dimensional In Vitro Models. Cells, 10.","DOI":"10.3390\/cells10010141"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"923","DOI":"10.1016\/j.tibtech.2018.03.011","article-title":"Inflammation-on-a-Chip: Probing the Immune System Ex Vivo","volume":"36","author":"Irimia","year":"2018","journal-title":"Trends Biotechnol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1239","DOI":"10.1111\/j.1582-4934.2011.01258.x","article-title":"Why is the partial oxygen pressure of human tissues a crucial parameter? Small molecules and hypoxia","volume":"15","author":"Carreau","year":"2011","journal-title":"J. Cell. Mol. Med."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"242","DOI":"10.1016\/j.stem.2009.08.009","article-title":"PO2 Matters in Stem Cell Culture","volume":"5","author":"Wion","year":"2009","journal-title":"Cell Stem Cell"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Kaneko, S., and Takamatsu, K. (2012). Cell Handling and Culture Under Controlled Oxygen Concentration. Biomedical Tissue Culture, InTech.","DOI":"10.5772\/52112"},{"key":"ref_10","first-page":"518","article-title":"Formation of Neutrophil Extracellular Traps under Low Oxygen Level","volume":"7","author":"Husein","year":"2016","journal-title":"Front. Immunol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"541","DOI":"10.3389\/fimmu.2017.00541","article-title":"Hypoxia Modulates the Response of Mast Cells to Staphylococcus aureus Infection","volume":"8","author":"Brogden","year":"2017","journal-title":"Front. Immunol."},{"key":"ref_12","first-page":"265","article-title":"Antimicrobial Activity of Mast Cells: Role and Relevance of Extracellular DNA Traps","volume":"7","year":"2016","journal-title":"Front. Immunol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"213","DOI":"10.1016\/j.neuro.2017.10.008","article-title":"The importance of controlling in vitro oxygen tension to accurately model in vivo neurophysiology","volume":"66","author":"Bordt","year":"2018","journal-title":"Neurotoxicology"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1016\/S1473-3099(07)70001-4","article-title":"Streptococcus suis: An emerging zoonotic pathogen","volume":"7","author":"Lun","year":"2007","journal-title":"Lancet Infect. Dis."},{"key":"ref_15","unstructured":"Locke, A.K., Zimmerman, J., Ramirez, A., Schwartz, K.J., and Stevenson, G. (2012). Streptococcosis. Diseases of Swine, Wiley-Blackwell."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1016\/j.vetmic.2003.09.018","article-title":"Epidemiology of Streptococcus suis serotype 5 infection in a pig herd with and without clinical disease","volume":"97","author":"Cloutier","year":"2003","journal-title":"Vet. Microbiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"371","DOI":"10.2217\/fmb.10.2","article-title":"Streptococcus suis: A new emerging or an old neglected zoonotic pathogen?","volume":"5","author":"Gottschalk","year":"2010","journal-title":"Future Microbiol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"617","DOI":"10.1086\/596763","article-title":"Streptococcus suis: An emerging human pathogen","volume":"48","author":"Wertheim","year":"2009","journal-title":"Clin. Infect. Dis."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"477","DOI":"10.4161\/viru.28595","article-title":"Streptococcus suis infection","volume":"5","author":"Feng","year":"2014","journal-title":"Virulence"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"423","DOI":"10.1016\/j.vetmic.2007.10.028","article-title":"Comparative evaluation of virulence and pathology of Streptococcus suis serotypes 2 and 9 in experimentally infected growers","volume":"128","author":"Beineke","year":"2008","journal-title":"Vet. Microbiol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1093\/infdis\/162.2.474","article-title":"Pathogenesis of meningitis caused by Streptococcus suis type 2","volume":"162","author":"Williams","year":"1990","journal-title":"J. Infect. Dis."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1038\/nri3391","article-title":"Orchestrated leukocyte recruitment to immune-privileged sites: Absolute barriers versus educational gates","volume":"13","author":"Shechter","year":"2013","journal-title":"Nat. Rev. Immunol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"4","DOI":"10.1186\/2045-8118-8-4","article-title":"Fluids and barriers of the CNS establish immune privilege by confining immune surveillance to a two-walled castle moat surrounding the CNS castle","volume":"8","author":"Engelhardt","year":"2011","journal-title":"Fluids Barriers CNS"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"39","DOI":"10.29245\/2572.942X\/2018\/4.1194","article-title":"Choroid Plexus Blood-CSF Barrier: Major Player in Brain Disease Modeling and Neuromedicine","volume":"3","author":"Johanson","year":"2018","journal-title":"J. Neurol. Neuromed."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"108478","DOI":"10.1016\/j.jneumeth.2019.108478","article-title":"Review of functional in vitro models of the blood-cerebrospinal fluid barrier in leukaemia research","volume":"329","author":"Erb","year":"2020","journal-title":"J. Neurosci. Methods"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1111\/j.1749-0774.2005.tb00059.x","article-title":"Establishment and characterization of a human malignant choroids plexus papilloma cell line (HIBCPP)","volume":"18","author":"Ishiwata","year":"2005","journal-title":"Hum. Cell"},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Schwerk, C., Papandreou, T., Schuhmann, D., Nickol, L., Borkowski, J., Steinmann, U., Quednau, N., Stump, C., Weiss, C., and Berger, J. (2012). Polar invasion and translocation of neisseria meningitidis and streptococcus suis in a novel human model of the blood-cerebrospinal fluid barrier. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0030069"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e12649","DOI":"10.1111\/cmi.12649","article-title":"Neutrophil extracellular trap formation in the Streptococcus suis-infected cerebrospinal fluid compartment","volume":"19","author":"Reuner","year":"2017","journal-title":"Cell. Microbiol."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1532","DOI":"10.1126\/science.1092385","article-title":"Neutrophil extracellular traps kill bacteria","volume":"303","author":"Brinkmann","year":"2004","journal-title":"Science"},{"key":"ref_30","doi-asserted-by":"crossref","unstructured":"de Buhr, N., Martens, A., Meurer, M., Bonilla, M.C., S\u00f6bbeler, F., Twele, L., Neudeck, S., Wendt, M., Beineke, A., and K\u00e4stner, S. (2021). In vivo oxygen measurement in cerebrospinal fluid of pigs to determine physiologic and pathophysiologic oxygen values during CNS infections. BMC Neurosci., 22.","DOI":"10.1186\/s12868-021-00648-x"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"125","DOI":"10.1016\/0378-1135(96)00028-4","article-title":"Characterization of virulence of the Sreptococcus suis serotype 2 reference strain Henrichsen S 735 in newborn gnotobiotic pigs","volume":"51","author":"Vecht","year":"1996","journal-title":"Vet. Microbiol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"1750","DOI":"10.1128\/IAI.67.4.1750-1756.1999","article-title":"Identification and characterization of the cps locus of Streptococcus suis serotype 2: The capsule protects against phagocytosis and is an important virulence factor","volume":"67","author":"Smith","year":"1999","journal-title":"Infect. Immun."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Meurer, M., \u00d6hlmann, S., Bonilla, M.C., Valentin-Weigand, P., Beineke, A., Hennig-Pauka, I., Schwerk, C., Schroten, H., Baums, C.G., and von K\u00f6ckritz-Blickwede, M. (2020). Role of Bacterial and Host DNases on Host-Pathogen Interaction during Streptococcus suis Meningitis. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21155289"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1007\/978-1-0716-0154-9_25","article-title":"Detection, Visualization, and Quantification of Neutrophil Extracellular Traps (NETs) and NET Markers","volume":"2087","year":"2020","journal-title":"Methods Mol. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1177\/2211068214561025","article-title":"TEER Measurement Techniques for In Vitro Barrier Model Systems","volume":"20","author":"Srinivasan","year":"2015","journal-title":"J. Lab. Autom."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1016\/j.freeradbiomed.2017.10.003","article-title":"Limitations of oxygen delivery to cells in culture: An underappreciated problem in basic and translational research","volume":"113","author":"Place","year":"2017","journal-title":"Free Radic. Biol. Med."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"66","DOI":"10.1016\/j.immuni.2013.11.020","article-title":"Transmigrating Neutrophils Shape the Mucosal Microenvironment through Localized Oxygen Depletion to Influence Resolution of Inflammation","volume":"40","author":"Campbell","year":"2014","journal-title":"Immunity"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"8238459","DOI":"10.1155\/2018\/8238459","article-title":"How Supraphysiological Oxygen Levels in Standard Cell Culture Affect Oxygen-Consuming Reactions","volume":"2018","author":"Stuart","year":"2018","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"5","DOI":"10.3389\/fnmol.2011.00005","article-title":"Oxygen, a Key Factor Regulating Cell Behavior during Neurogenesis and Cerebral Diseases","volume":"4","author":"Zhang","year":"2011","journal-title":"Front. Mol. Neurosci."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1186\/s13287-015-0124-z","article-title":"Do hypoxia\/normoxia culturing conditions change the neuroregulatory profile of Wharton Jelly mesenchymal stem cell secretome?","volume":"6","author":"Teixeira","year":"2015","journal-title":"Stem Cell Res. Ther."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Chadwick, W., Boyle, J.P., Zhou, Y., Wang, L., Park, S.-S., Martin, B., Wang, R., Becker, K.G., Wood, W.H., and Zhang, Y. (2011). Multiple Oxygen Tension Environments Reveal Diverse Patterns of Transcriptional Regulation in Primary Astrocytes. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0021638"},{"key":"ref_42","doi-asserted-by":"crossref","unstructured":"Al-Ani, A., Toms, D., Kondro, D., Thundathil, J., Yu, Y., and Ungrin, M. (2018). Oxygenation in cell culture: Critical parameters for reproducibility are routinely not reported. PLoS ONE, 13.","DOI":"10.1371\/journal.pone.0204269"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"e246","DOI":"10.1038\/cddis.2011.128","article-title":"Oxygen matters: Tissue culture oxygen levels affect mitochondrial function and structure as well as responses to HIV viroproteins","volume":"2","author":"Tiede","year":"2011","journal-title":"Cell Death Dis."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Krzywinska, E., and Stockmann, C. (2018). Hypoxia, Metabolism and Immune Cell Function. Biomedicines, 6.","DOI":"10.3390\/biomedicines6020056"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1186\/1742-2094-10-31","article-title":"Transmigration of polymorphnuclear neutrophils and monocytes through the human blood-cerebrospinal fluid barrier after bacterial infection in vitro","volume":"10","author":"Steinmann","year":"2013","journal-title":"J. Neuroinflamm."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1742","DOI":"10.1128\/iai.62.5.1742-1748.1994","article-title":"Identification, purification, and characterization of a thiol-activated hemolysin (suilysin) of Streptococcus suis","volume":"62","author":"Jacobs","year":"1994","journal-title":"Infect. Immun."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.femsim.2004.12.006","article-title":"Strain-dependent disruption of blood-cerebrospinal fluid barrier by Streptoccocus suis in vitro","volume":"44","author":"Tenenbaum","year":"2005","journal-title":"FEMS Immunol. Med. Microbiol."},{"key":"ref_48","doi-asserted-by":"crossref","unstructured":"Bonilla, M.C., Fingerhut, L., Alfonso-Castro, A., Mergani, A., Schwennen, C., von K\u00f6ckritz-Blickwede, M., and de Buhr, N. (2020). How Long Does a Neutrophil Live?\u2014The Effect of 24 h Whole Blood Storage on Neutrophil Functions in Pigs. Biomedicines, 8.","DOI":"10.3390\/biomedicines8080278"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"3431","DOI":"10.1182\/blood-2016-03-700336","article-title":"Human neutrophil kinetics: Modeling of stable isotope labeling data supports short blood neutrophil half-lives","volume":"127","author":"Elemans","year":"2016","journal-title":"Blood"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/1\/151\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,13]],"date-time":"2025-10-13T14:12:35Z","timestamp":1760364755000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/11\/1\/151"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,1,4]]},"references-count":49,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2022,1]]}},"alternative-id":["cells11010151"],"URL":"https:\/\/doi.org\/10.3390\/cells11010151","relation":{},"ISSN":["2073-4409"],"issn-type":[{"type":"electronic","value":"2073-4409"}],"subject":[],"published":{"date-parts":[[2022,1,4]]}}}