{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,13]],"date-time":"2026-05-13T20:23:13Z","timestamp":1778703793273,"version":"3.51.4"},"reference-count":48,"publisher":"MDPI AG","issue":"9","license":[{"start":{"date-parts":[[2021,4,26]],"date-time":"2021-04-26T00:00:00Z","timestamp":1619395200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>We have developed a membrane filter-assisted cell-based biosensing platform by using a polyester membrane as a three-dimensional (3D) cell culture scaffold in which cells can be grown by physical attachment. The membrane was simply treated with ethanol to increase surficial hydrophobicity, inducing the stable settlement of cells via gravity. The 3D membrane scaffold was able to provide a relatively longer cell incubation time (up to 16 days) as compared to a common two-dimensional (2D) cell culture environment. For a practical application, we fabricated a cylindrical cartridge to support the scaffold membranes stacked inside the cartridge, enabling not only the maintenance of a certain volume of culture media but also the simple exchange of media in a flow-through manner. The cartridge-type cell-based analytical system was exemplified for pathogen detection by measuring the quantities of toll-like receptor 1 (TLR1) induced by applying a lysate of P. aeruginosa and live E. coli, respectively, providing a fast, convenient colorimetric TLR1 immunoassay. The color images of membranes were digitized to obtain the response signals. We expect the method to further be applied as an alternative tool to animal testing in various research areas such as cosmetic toxicity and drug efficiency.<\/jats:p>","DOI":"10.3390\/s21093042","type":"journal-article","created":{"date-parts":[[2021,4,27]],"date-time":"2021-04-27T06:19:11Z","timestamp":1619504351000},"page":"3042","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["A Membrane Filter-Assisted Mammalian Cell-Based Biosensor Enabling 3D Culture and Pathogen Detection"],"prefix":"10.3390","volume":"21","author":[{"given":"Il-Hoon","family":"Cho","sequence":"first","affiliation":[{"name":"BK21 Plus Program, Department of Senior Healthcare, Graduate School, Eulji University, Daejeon 34824, Korea"},{"name":"Department of Biomedical Laboratory Science, College of Health Science, Eulji University, 553 Sanseong-Daero, Sujeong-gu, Seongnam 13135, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jin-Woo","family":"Jeon","sequence":"additional","affiliation":[{"name":"Department of Bio-Microsystem Technology, Korea University, Seoul 02841, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Min-Ji","family":"Choi","sequence":"additional","affiliation":[{"name":"BK21 Plus Program, Department of Senior Healthcare, Graduate School, Eulji University, Daejeon 34824, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Hyun-Mo","family":"Cho","sequence":"additional","affiliation":[{"name":"Division of Interdisciplinary Materials Measurement Institute, Korea Research Institute of Standards and Science, Daejeon 34113, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5073-2619","authenticated-orcid":false,"given":"Jong-Sung","family":"Lee","sequence":"additional","affiliation":[{"name":"Department of Genetic Engineering, College of Biotechnology and Bioengineering, Sungkyunkwan University, Suwon 16419, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Dong-Hyung","family":"Kim","sequence":"additional","affiliation":[{"name":"Division of Interdisciplinary Materials Measurement Institute, Korea Research Institute of Standards and Science, Daejeon 34113, Korea"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2021,4,26]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"179","DOI":"10.1016\/j.tibtech.2008.11.006","article-title":"Mammalian cell-based biosensors for pathogens and toxins","volume":"27","author":"Banerjee","year":"2009","journal-title":"Trends Biotechnol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"576","DOI":"10.1016\/j.snb.2004.11.056","article-title":"Cell-based biosensors and its application in biomedicine","volume":"108","author":"Wang","year":"2005","journal-title":"Sens. Actuators B Chem."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"6423","DOI":"10.1021\/cr2003129","article-title":"Cell-Based Biosensors and Their Application in Biomedicine","volume":"114","author":"Liu","year":"2014","journal-title":"Chem. Rev."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"159","DOI":"10.1124\/jpet.108.149724","article-title":"Fluorescent Ligands of the Bradykinin B1 Receptors: Pharmacologic Characterization and Application to the Study of Agonist-Induced Receptor Translocation and Cell Surface Receptor Expression","volume":"329","author":"Bawolak","year":"2009","journal-title":"J. Pharmacol. Exp. Ther."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1038\/nmeth1079","article-title":"MicroRNA sponges: Competitive inhibitors of small RNAs in mammalian cells","volume":"4","author":"Ebert","year":"2007","journal-title":"Nat. Methods"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"9713","DOI":"10.1021\/ac300668y","article-title":"Toll-Like Receptor-Based Immuno-Analysis of Pathogenic Microorganisms","volume":"84","author":"Cho","year":"2012","journal-title":"Anal. Chem."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"135","DOI":"10.1007\/s10544-006-9006-6","article-title":"Screening cell surface receptors using micromosaic immunoassays","volume":"9","author":"Wolf","year":"2007","journal-title":"Biomed. Microdevices"},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Kohl, T.O., and Ascoli, C.A. (2017). Direct and Indirect Cell-Based Enzyme-Linked Immunosorbent Assay. Cold Spring Harb. Protoc., 2017.","DOI":"10.1101\/pdb.prot093732"},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Lu, X., Ye, Y., Zhang, Y., and Sun, X. (2020). Current research progress of mammalian cell-based biosensors on the detection of foodborne pathogens and toxins. Crit. Rev. Food Sci. Nutr.","DOI":"10.1080\/10408398.2020.1809341"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1016\/S1367-5931(00)00110-1","article-title":"High-throughput screening: New technology for the 21st century","volume":"4","author":"Hertzberg","year":"2000","journal-title":"Curr. Opin. Chem. Biol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1153","DOI":"10.1016\/j.bios.2008.06.060","article-title":"An impedimetric biosensor based on PC 12 cells for the monitoring of exogenous agents","volume":"24","author":"Slaughter","year":"2009","journal-title":"Biosens. Bioelectron."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"879","DOI":"10.1039\/b800394g","article-title":"Drug testing on 3D in vitro tissues trapped on a microcavity chip","volume":"8","author":"Kloss","year":"2008","journal-title":"Lab Chip"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1757","DOI":"10.1016\/j.bios.2004.06.037","article-title":"Cell-based biosensors based on light-addressable potentiometric sensors for single cell monitoring","volume":"20","author":"Xu","year":"2005","journal-title":"Biosens. Bioelectron."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1016\/j.aca.2016.04.011","article-title":"Development of a mediated whole cell-based electrochemical biosensor for joint toxicity assessment of multi-pollutants using a mixed microbial consortium","volume":"924","author":"Gao","year":"2016","journal-title":"Anal. Chim. Acta"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"126","DOI":"10.1073\/pnas.60.1.126","article-title":"Growth control in cultured cells: Selection of sublines with increased sensitivity to contact inhibition and decreased tumor-producing ability","volume":"60","author":"Pollack","year":"1968","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"325","DOI":"10.1002\/jcp.1030660310","article-title":"The initiation of cell division in a contact-inhibited mammalian cell line","volume":"66","author":"Todaro","year":"1965","journal-title":"J. Cell. Comp. Physiol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"264","DOI":"10.1016\/S0962-8924(03)00057-6","article-title":"Fibroblast biology in three-dimensional collagen matrices","volume":"13","author":"Grinnell","year":"2003","journal-title":"Trends Cell Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"384","DOI":"10.1091\/mbc.e02-08-0493","article-title":"Dendritic Fibroblasts in Three-dimensional Collagen Matrices","volume":"14","author":"Grinnell","year":"2003","journal-title":"Mol. Biol. Cell"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"8228","DOI":"10.1016\/j.biomaterials.2010.07.035","article-title":"Patterning network structure to spatially control cellular remodeling and stem cell fate within 3-dimensional hydrogels","volume":"31","author":"Khetan","year":"2010","journal-title":"Biomaterials"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"2962","DOI":"10.1016\/j.biomaterials.2008.04.004","article-title":"Three-dimensional micropatterning of bioactive hydrogels via two-photon laser scanning photolithography for guided 3D cell migration","volume":"29","author":"Lee","year":"2008","journal-title":"Biomaterials"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"2","DOI":"10.1016\/j.matbio.2006.09.008","article-title":"On the origins of the extracellular matrix in vertebrates","volume":"26","author":"Robertson","year":"2007","journal-title":"Matrix Biol."},{"key":"ref_22","first-page":"3015","article-title":"Deconstructing the third dimension\u2014how 3D culture microenvironments alter cellular cues","volume":"125","author":"Baker","year":"2012","journal-title":"J. Cell Sci."},{"key":"ref_23","unstructured":"Rahman, M. (2012). Degradation of Polyesters in Medical Applications, IntechOpen."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"5020","DOI":"10.1021\/acsanm.9b00956","article-title":"Quantifying the Numbers of Gold Nanoparticles in the Test Zone of Lateral Flow Immunoassay Strips","volume":"2","author":"Khlebtsov","year":"2019","journal-title":"ACS Appl. Nano Mater."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"111","DOI":"10.1042\/EBC20150012","article-title":"Lateral flow assays","volume":"60","author":"Koczula","year":"2016","journal-title":"Essays Biochem."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"024117","DOI":"10.1063\/1.5021002","article-title":"A high-performance polydimethylsiloxane electrospun membrane for cell culture in lab-on-a-chip","volume":"12","author":"Moghadas","year":"2018","journal-title":"Biomicrofluidics"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1016\/j.bios.2019.01.050","article-title":"A novel electrochemical mast cell-based paper biosensor for the rapid detection of milk allergen casein","volume":"130","author":"Jiang","year":"2019","journal-title":"Biosens. Bioelectron."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"32","DOI":"10.1016\/j.mattod.2016.07.001","article-title":"Paper-based cell culture platform and its emerging biomedical applications","volume":"20","author":"Ng","year":"2017","journal-title":"Mater. Today"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"138","DOI":"10.1007\/s00284-011-9959-4","article-title":"Up-regulation of bradykinin B2 receptor by Pseudomonas aeruginosa via the NF-\u03baB pathway","volume":"63","author":"Shin","year":"2011","journal-title":"Curr. Microbiol."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"302","DOI":"10.1006\/abio.1996.0171","article-title":"Linearization of the Bradford Protein Assay Increases Its Sensitivity: Theoretical and Experimental Studies","volume":"236","author":"Zor","year":"1996","journal-title":"Anal. Biochem."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1417","DOI":"10.1093\/jnci\/51.5.1417","article-title":"In Vitro Cultivation of Human Tumors: Establishment of Cell Lines Derived From a Series of Solid Tumors2","volume":"51","author":"Giard","year":"1973","journal-title":"J. Natl. Cancer Inst."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Jeon, J.-W., Ha, U.-H., and Paek, S.-H. (2014). In Vitro Inflammation Inhibition Model Based on Semi-Continuous Toll-Like Receptor Biosensing. PLoS ONE, 9.","DOI":"10.1371\/journal.pone.0105212"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1083\/jcb.136.1.1","article-title":"Anchorage-dependent Cell Cycle Progression","volume":"136","author":"Assoian","year":"1997","journal-title":"J. Cell Biol."},{"key":"ref_34","doi-asserted-by":"crossref","unstructured":"Chen, M., Huang, J., Yang, X., Liu, B., Zhang, W., Huang, L., Deng, F., Ma, J., Bai, Y., and Lu, R. (2012). Serum Starvation Induced Cell Cycle Synchronization Facilitates Human Somatic Cells Reprogramming. PLoS ONE, 7.","DOI":"10.1371\/journal.pone.0028203"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"032204","DOI":"10.1063\/1.3466882","article-title":"Oxygen plasma treatment for reducing hydrophobicity of a sealed polydimethylsiloxane microchannel","volume":"4","author":"Tan","year":"2010","journal-title":"Biomicrofluidics"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"170","DOI":"10.1016\/j.snb.2005.04.037","article-title":"Thermal aging and reduced hydrophobic recovery of polydimethylsiloxane","volume":"114","author":"Eddington","year":"2006","journal-title":"Sens. Actuators B Chem."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"9971","DOI":"10.1021\/la0205553","article-title":"Chemical Modification of the Surface of Poly(dimethylsiloxane) by Atom-Transfer Radical Polymerization of Acrylamide","volume":"18","author":"Xiao","year":"2002","journal-title":"Langmuir"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"476","DOI":"10.1038\/196476a0","article-title":"Effect of absorbed proteins on the wettability of hydrophilic and hydrophobic solids","volume":"196","author":"Vroman","year":"1962","journal-title":"Nature"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1016\/S1381-5148(02)00086-X","article-title":"Esterification of maleic acid with ethanol over cation-exchange resin catalysts","volume":"52","author":"Yadav","year":"2002","journal-title":"React. Funct. Polym."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"2333","DOI":"10.1016\/S0142-9612(99)00162-3","article-title":"Surface micropatterning to regulate cell functions","volume":"20","author":"Ito","year":"1999","journal-title":"Biomaterials"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Da Cunha, C.B., Lopes, P.P., Mayer, F.D., and Hoffmann, R. (2018). Assessment of Chemical and Mechanical Properties of Polymers Aiming to Replace the Stainless Steel in Distillation Column. Mater. Res., 21.","DOI":"10.1590\/1980-5373-mr-2017-0679"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"519","DOI":"10.1016\/S0165-2370(03)00024-X","article-title":"Thermal degradation of poly(vinyl chloride)","volume":"70","author":"Marongiu","year":"2003","journal-title":"J. Anal. Appl. Pyrolysis"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"19","DOI":"10.1016\/j.bios.2016.04.027","article-title":"A fluorescent immunosensor for high-sensitivity cardiac troponin I using a spatially-controlled polymeric, nano-scale tracer to prevent quenching","volume":"83","author":"Seo","year":"2016","journal-title":"Biosens. Bioelectron."},{"key":"ref_44","first-page":"663","article-title":"The morphometrical analysis on the ultrastructure of A549 cells","volume":"51","author":"Jiang","year":"2010","journal-title":"Rom. J. Morphol. Embryol."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"74","DOI":"10.1006\/abio.1999.4309","article-title":"Colorimetric Determination of Cell Numbers by Janus Green Staining","volume":"275","author":"Raspotnig","year":"1999","journal-title":"Anal. Biochem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"293","DOI":"10.1038\/nrd3203","article-title":"Targeting Toll-like receptors: Emerging therapeutics?","volume":"9","author":"Hennessy","year":"2010","journal-title":"Nat. Rev. Drug Discov."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1016\/0142-9612(95)93257-E","article-title":"Mechanism of cell detachment from temperature-modulated, hydrophilic-hydrophobic polymer surfaces","volume":"16","author":"Okano","year":"1995","journal-title":"Biomaterials"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1002\/jcp.1040700104","article-title":"Dynamic adhesion and separation of cellsin vitro II. Interactions of cells with hydrophilic and hydrophobic surfaces","volume":"70","author":"Weiss","year":"1967","journal-title":"J. Cell. Physiol."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/21\/9\/3042\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T05:53:04Z","timestamp":1760161984000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/21\/9\/3042"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2021,4,26]]},"references-count":48,"journal-issue":{"issue":"9","published-online":{"date-parts":[[2021,5]]}},"alternative-id":["s21093042"],"URL":"https:\/\/doi.org\/10.3390\/s21093042","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,4,26]]}}}