{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,17]],"date-time":"2026-07-17T15:33:05Z","timestamp":1784302385196,"version":"3.55.0"},"reference-count":20,"publisher":"MDPI AG","issue":"7","license":[{"start":{"date-parts":[[2023,3,28]],"date-time":"2023-03-28T00:00:00Z","timestamp":1679961600000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Epithelial cells may play an important role in the pathologic process of chronic rhinosinusitis with nasal polyps. Therefore, providing epithelial cells from a biobank could greatly contribute to further research. In the present work, the isolation of epithelial cells from long-term cryopreserved tissue is demonstrated. Polyp tissues were cryopreserved in a commercially available freezing medium with dimethyl sulfoxide and stored in liquid nitrogen. The outgrowth and proliferation of epithelial cells from cryopreserved tissue were evaluated and compared to that of fresh tissue. Flow cytometric analysis with anti-cytokeratin, anti-p63, and anti-Ki-67 was performed to identify epithelial cells and determine differentiation and proliferation. A functionality test was performed by determining type 2\u2013relevant proteins, representatively thymic stromal lymphopoietin (TSLP) and periostin, using ELISA. Primary epithelial cells could be isolated from cryopreserved tissues. Cells from cryopreserved tissues showed comparable outgrowth and proliferation to that of fresh tissue. Isolated epithelial cells showed high cytokeratin, p63, and Ki-67 expression and secreted TSLP and periostin. In the present study, a method for long-term cryopreservation of polyp tissue was established, thereby enabling the isolation and cell culture of primary cell culture at a later time. Epithelial cell availability should be greatly improved by including this method in a biobank.<\/jats:p>","DOI":"10.3390\/ijms24076383","type":"journal-article","created":{"date-parts":[[2023,3,29]],"date-time":"2023-03-29T02:03:53Z","timestamp":1680055433000},"page":"6383","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":2,"title":["Long-Term Cryopreservation of Nasal Polyp Tissue in a Biobank for the Isolation and Culture of Primary Epithelial Cells"],"prefix":"10.3390","volume":"24","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3107-7951","authenticated-orcid":false,"given":"Jonghui","family":"Kim","sequence":"first","affiliation":[{"name":"Department of Oto-Rhino-Laryngology\u2014Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten\/Herdecke, D-58097 Hagen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2165-1913","authenticated-orcid":false,"given":"Karla","family":"Hegener","sequence":"additional","affiliation":[{"name":"Department of Oto-Rhino-Laryngology\u2014Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten\/Herdecke, D-58097 Hagen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7010-4807","authenticated-orcid":false,"given":"Claudia","family":"Hagedorn","sequence":"additional","affiliation":[{"name":"Chair of Biochemistry and Molecular Medicine, Center for Biomedical Education and Research, School of Life Sciences (ZBAF), University of Witten\/Herdecke, D-58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-6989-8694","authenticated-orcid":false,"given":"Kaschin","family":"Jamal Jameel","sequence":"additional","affiliation":[{"name":"Medical Clinic III for Pneumology, Allergology and Sleep Medicine, Bergmannsheil University Hospital, Ruhr-University Bochum, D-44789 Bochum, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-1468-0273","authenticated-orcid":false,"given":"Daniel","family":"Weidinger","sequence":"additional","affiliation":[{"name":"Medical Clinic III for Pneumology, Allergology and Sleep Medicine, Bergmannsheil University Hospital, Ruhr-University Bochum, D-44789 Bochum, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0295-5447","authenticated-orcid":false,"given":"Inga Marte Charlott","family":"Seuthe","sequence":"additional","affiliation":[{"name":"Department of Oto-Rhino-Laryngology\u2014Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten\/Herdecke, D-58097 Hagen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7877-7977","authenticated-orcid":false,"given":"Sabine","family":"Eichhorn","sequence":"additional","affiliation":[{"name":"Department of Oto-Rhino-Laryngology\u2014Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten\/Herdecke, D-58097 Hagen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0213-7815","authenticated-orcid":false,"given":"Florian","family":"Kreppel","sequence":"additional","affiliation":[{"name":"Chair of Biochemistry and Molecular Medicine, Center for Biomedical Education and Research, School of Life Sciences (ZBAF), University of Witten\/Herdecke, D-58453 Witten, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6868-0055","authenticated-orcid":false,"given":"J\u00fcrgen","family":"Knobloch","sequence":"additional","affiliation":[{"name":"Medical Clinic III for Pneumology, Allergology and Sleep Medicine, Bergmannsheil University Hospital, Ruhr-University Bochum, D-44789 Bochum, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4582-819X","authenticated-orcid":false,"given":"Jonas Jae-Hyun","family":"Park","sequence":"additional","affiliation":[{"name":"Department of Oto-Rhino-Laryngology\u2014Head and Neck Surgery, St.-Josefs-Hospital Hagen, University of Witten\/Herdecke, D-58097 Hagen, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2023,3,28]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"86","DOI":"10.1038\/s41572-020-00218-1","article-title":"Adult Chronic Rhinosinusitis","volume":"6","author":"Bachert","year":"2020","journal-title":"Nat. Rev. Dis. Prim."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"1449","DOI":"10.1016\/j.jaci.2015.12.1324","article-title":"Inflammatory Endotypes of Chronic Rhinosinusitis Based on Cluster Analysis of Biomarkers","volume":"137","author":"Tomassen","year":"2016","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1638","DOI":"10.1016\/S0140-6736(19)31881-1","article-title":"Efficacy and Safety of Dupilumab in Patients with Severe Chronic Rhinosinusitis with Nasal Polyps (LIBERTY NP SINUS-24 and LIBERTY NP SINUS-52): Results from Two Multicentre, Randomised, Double-Blind, Placebo-Controlled, Parallel-Group Phase 3 Trials","volume":"394","author":"Bachert","year":"2019","journal-title":"Lancet"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"595","DOI":"10.1016\/j.jaci.2020.05.032","article-title":"Efficacy and Safety of Omalizumab in Nasal Polyposis: 2 Randomized Phase 3 Trials","volume":"146","author":"Gevaert","year":"2020","journal-title":"J. Allergy Clin. Immunol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1141","DOI":"10.1016\/S2213-2600(21)00097-7","article-title":"Mepolizumab for Chronic Rhinosinusitis with Nasal Polyps (SYNAPSE): A Randomised, Double-Blind, Placebo-Controlled, Phase 3 Trial","volume":"9","author":"Han","year":"2021","journal-title":"Lancet Respir. Med."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"432","DOI":"10.1164\/rccm.201212-2227OC","article-title":"IL-33\u2013Responsive Innate Lymphoid Cells Are an Important Source of IL-13 in Chronic Rhinosinusitis with Nasal Polyps","volume":"188","author":"Shaw","year":"2013","journal-title":"Am. J. Respir. Crit. Care Med."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"186","DOI":"10.4168\/aair.2011.3.3.186","article-title":"Increased Expression and Role of Thymic Stromal Lymphopoietin in Nasal Polyposis","volume":"3","author":"Kimura","year":"2011","journal-title":"Allergy Asthma Immunol. Res."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"333","DOI":"10.1016\/j.anai.2020.01.018","article-title":"Epithelial Cell Function and Remodeling in Nasal Polyposis","volume":"124","author":"Bankova","year":"2020","journal-title":"Ann. Allergy Asthma Immunol."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Chiarella, E., Lombardo, N., Lobello, N., Aloisio, A., Aragona, T., Pelaia, C., Scicchitano, S., Bond, H.M., and Mesuraca, M. (2020). Nasal Polyposis: Insights in Epithelial-Mesenchymal Transition and Differentiation of Polyp Mesenchymal Stem Cells. Int. J. Mol. Sci., 21.","DOI":"10.20944\/preprints202008.0087.v1"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"649","DOI":"10.1038\/s41586-018-0449-8","article-title":"Allergic Inflammatory Memory in Human Respiratory Epithelial Progenitor Cells","volume":"560","author":"Dwyer","year":"2018","journal-title":"Nature"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"327","DOI":"10.3109\/01902148.2014.925987","article-title":"Comparison of Three Different Brushing Techniques to Isolate and Culture Primary Nasal Epithelial Cells from Human Subjects","volume":"40","author":"Stokes","year":"2014","journal-title":"Exp. Lung Res."},{"key":"ref_12","first-page":"183","article-title":"Well-Differentiated Human Airway Epithelial Cell Cultures","volume":"107","author":"Fulcher","year":"2005","journal-title":"Methods Mol. Med."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"100782","DOI":"10.1016\/j.xpro.2021.100782","article-title":"Isolation, Expansion, Differentiation, and Histological Processing of Human Nasal Epithelial Cells","volume":"2","author":"Manna","year":"2021","journal-title":"STAR Protoc."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"296","DOI":"10.3109\/00016489.2013.859396","article-title":"Isolation and Culture of Primary Human Nasal Epithelial Cells from Anesthetized Nasal Epithelia","volume":"134","author":"Hussain","year":"2014","journal-title":"Acta Otolaryngol."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Kim, J., Hegener, K., Hagedorn, C., Jamal Jameel, K., Weidinger, D., Seuthe, I.M.C., Eichhorn, S., Kreppel, F., Park, J.J.-H., and Knobloch, J. (2023). Simple, Low-Cost, and Well-Performing Method, the Outgrowth Technique, for the Isolation of Epithelial Cells from Nasal Polyps. bioRxiv, 2023.01.10.522992.","DOI":"10.1101\/2023.01.10.522992"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1369\/0022155419892965","article-title":"MiR-210 Is Overexpressed in Tumor-Infiltrating Plasma Cells in Triple-Negative Breast Cancer","volume":"68","author":"Bar","year":"2020","journal-title":"J. Histochem. Cytochem."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Kim, T.-H., Lee, J.-Y., Park, J.-S., Park, S.-W., Jang, A.-S., Lee, J.-Y., Byun, J.-Y., Uh, S.-T., Koh, E.-S., and Chung, I.Y. (2011). Fatty Acid Binding Protein 1 Is Related with Development of Aspirin-Exacerbated Respiratory Disease. PLoS ONE, 6.","DOI":"10.1371\/journal.pone.0022711"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"112","DOI":"10.1007\/s10875-010-9462-x","article-title":"KIF3A, a Cilia Structural Gene on Chromosome 5q31, and Its Polymorphisms Show an Association with Aspirin Hypersensitivity in Asthma","volume":"31","author":"Kim","year":"2011","journal-title":"J. Clin. Immunol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1038\/s41588-018-0314-6","article-title":"A Loss-of-Function Variant in ALOX15 Protects against Nasal Polyps and Chronic Rhinosinusitis","volume":"51","author":"Kristjansson","year":"2019","journal-title":"Nat. Genet."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Coles, J.L., Thompson, J., Horton, K.L., Hirst, R.A., Griffin, P., Williams, G.M., Goggin, P., Doherty, R., Lackie, P.M., and Harris, A. (2020). A Revised Protocol for Culture of Airway Epithelial Cells as a Diagnostic Tool for Primary Ciliary Dyskinesia. J. Clin. Med., 9.","DOI":"10.3390\/jcm9113753"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/7\/6383\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T19:05:13Z","timestamp":1760123113000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/7\/6383"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,3,28]]},"references-count":20,"journal-issue":{"issue":"7","published-online":{"date-parts":[[2023,4]]}},"alternative-id":["ijms24076383"],"URL":"https:\/\/doi.org\/10.3390\/ijms24076383","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,3,28]]}}}