{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,24]],"date-time":"2026-01-24T10:27:45Z","timestamp":1769250465922,"version":"3.49.0"},"reference-count":31,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2023,9,30]],"date-time":"2023-09-30T00:00:00Z","timestamp":1696032000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100004329","name":"Slovenian Research Agency","doi-asserted-by":"publisher","award":["P3-0289"],"award-info":[{"award-number":["P3-0289"]}],"id":[{"id":"10.13039\/501100004329","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Cells"],"abstract":"<jats:p>Tumor spheroids in the ascites of high-grade serous carcinoma (HGSC) are poorly described. Our objective was to describe their morphological features, cellular composition, PD-1 and PD-L1 expression, and survival correlation of these parameters. The density and size of spheroids were assessed in Giemsa-stained smears; the cell composition of spheroids, including tumor cells, immune cells, capillaries, and myofibroblasts, as well as PD-1 and PD-L1 expression on tumor and immune cells was assessed in immunocytochemically stained cell block sections. Forty-seven patients with primary HGSC and malignant ascites were included. A cut-off value for a spheroid density of 10% was established, which significantly predicted overall survival. However, spheroid size did not correlate with survival outcomes. Spheroids were primarily composed of tumor cells, but the presence of lymphocytes and macrophages was also confirmed. Moreover, capillaries were present in the spheroids of three patients, but the presence of myofibroblasts was not confirmed. PD-1 was expressed on lymphocytes but not on tumor cells. PD-L1 expression was seen on both tumor and immune cells, assessed by 22C3 and SP263 antibody clones but not by the SP142 clone. Our results highlight the potential of routine cytopathological techniques to analyze spheroids in HGSC ascites as a valuable tool to investigate their potential as prognostic markers.<\/jats:p>","DOI":"10.3390\/cells12192390","type":"journal-article","created":{"date-parts":[[2023,10,1]],"date-time":"2023-10-01T16:58:09Z","timestamp":1696179489000},"page":"2390","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Morphological and Immunocytochemical Characterization of Tumor Spheroids in Ascites from High-Grade Serous Carcinoma"],"prefix":"10.3390","volume":"12","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1889-1991","authenticated-orcid":false,"given":"Simona","family":"Miceska","sequence":"first","affiliation":[{"name":"Department of Cytopathology, Institute of Oncology Ljubljana, Zalo\u0161ka Cesta 2, 1000 Ljubljana, Slovenia"},{"name":"Faculty of Medicine, University of Ljubljana, Korytkova Ulica 2, 1000 Ljubljana, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7076-4186","authenticated-orcid":false,"given":"Erik","family":"\u0160kof","sequence":"additional","affiliation":[{"name":"Faculty of Medicine, University of Ljubljana, Korytkova Ulica 2, 1000 Ljubljana, Slovenia"},{"name":"Department of Medical Oncology, Institute of Oncology Ljubljana, Zalo\u0161ka Cesta 2, 1000 Ljubljana, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8332-3627","authenticated-orcid":false,"given":"Gorana","family":"Ga\u0161ljevi\u0107","sequence":"additional","affiliation":[{"name":"Department of Pathology, Institute of Oncology Ljubljana, Zalo\u0161ka Cesta 2, 1000 Ljubljana, Slovenia"},{"name":"Faculty of Medicine, University of Maribor, Taborska Ulica 8, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-0605-6596","authenticated-orcid":false,"given":"Veronika","family":"Kloboves-Prevodnik","sequence":"additional","affiliation":[{"name":"Department of Cytopathology, Institute of Oncology Ljubljana, Zalo\u0161ka Cesta 2, 1000 Ljubljana, Slovenia"},{"name":"Faculty of Medicine, University of Maribor, Taborska Ulica 8, 2000 Maribor, Slovenia"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,9,30]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"284","DOI":"10.3322\/caac.21456","article-title":"Ovarian cancer statistics","volume":"68","author":"Torre","year":"2018","journal-title":"CA Cancer J. Clin."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"15818","DOI":"10.18632\/oncotarget.24530","article-title":"WT1, p53 and p16 expression in the diagnosis of low- and high-grade serous ovarian carcinomas and their relation to prognosis","volume":"9","author":"Sallum","year":"2018","journal-title":"Oncotarget"},{"key":"ref_3","doi-asserted-by":"crossref","unstructured":"Rakina, M., Kazakova, A., Villert, A., Kolomiets, L., and Larionova, I. (2022). Spheroid formation and peritoneal metastasis in ovarian cancer: The role of stromal and immune components. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23116215"},{"key":"ref_4","first-page":"4","article-title":"Disaggregation and invasion of ovarian carcinoma ascites spheroids","volume":"24","author":"Burleson","year":"2006","journal-title":"J. Transl. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"717","DOI":"10.1016\/j.stemcr.2020.03.004","article-title":"Developing Organoids from Ovarian Cancer as Experimental and Preclinical Models","volume":"14","author":"Maenhoudt","year":"2020","journal-title":"Stem Cell Rep."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"614","DOI":"10.1038\/s41418-021-00878-w","article-title":"Single cell-derived spheroids capture the self-renewing subpopulations of metastatic ovarian cancer","volume":"29","author":"Velletri","year":"2022","journal-title":"Cell Death Differ."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"188","DOI":"10.1002\/cncy.22664","article-title":"Cytopathological assessment is an accurate method for identifying immunophenotypic features and BRCA1\/2 mutations of high-grade serous carcinoma from ascites","volume":"131","author":"Miceska","year":"2023","journal-title":"Cancer Cytopathol."},{"key":"ref_8","doi-asserted-by":"crossref","unstructured":"Micek, H.M., Rosenstock, L., Ma, Y., Hielsberg, C., Montemorano, L., Gari, M.K., Ponik, S.M., and Kreeger, P.K. (2023). Model of collective detachment in high-grade serous ovarian cancer demonstrates that tumor spheroids produce ECM to support metastatic processes. APL Bioeng., 7.","DOI":"10.1063\/5.0132254"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1080\/15384047.2015.1040960","article-title":"Tumor infiltrating lymphocytes in ovarian cancer","volume":"16","author":"Santoiemma","year":"2015","journal-title":"Cancer Biol. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1111","DOI":"10.1080\/21645515.2019.1571892","article-title":"PD-1 and PD-L1 in cancer immunotherapy: Clinical implications and future considerations","volume":"15","author":"Jiang","year":"2019","journal-title":"Hum. Vaccines Immunother."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1389","DOI":"10.1136\/ijgc-2019-000609","article-title":"Prognostic significance of PD-1 and PD-L1 positive tumor-infiltrating immune cells in ovarian carcinoma","volume":"29","author":"Buderath","year":"2019","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1271","DOI":"10.21037\/tcr.2018.10.11","article-title":"Characterization of PD-L1 and its prognostic value in patients with ovarian cancer","volume":"7","author":"Cheng","year":"2018","journal-title":"Transl. Cancer Res."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"3750","DOI":"10.1038\/s41598-021-83276-z","article-title":"Differential expression of PD-L1 between primary and metastatic epithelial ovarian cancer and its clinico-pathological correlation","volume":"11","author":"Parvathareddy","year":"2021","journal-title":"Sci. Rep."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"607","DOI":"10.1016\/j.ygyno.2016.12.021","article-title":"Tumor-associated macrophage expression of PD-L1 in implants of high grade serous ovarian carcinoma: A comparison of matched primary and metastatic tumors","volume":"144","author":"Gottlieb","year":"2017","journal-title":"Gynecol. Oncol."},{"key":"ref_15","unstructured":"Miceska, S. (2023). Immune Cells in Effusions in Patients with High Grade Serous Ovarian Cancer. [Ph.D. Thesis, University of Ljubljana]."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"140","DOI":"10.1089\/adt.2018.895","article-title":"Quantitative Size-Based Analysis of Tumor Spheroids and Responses to Therapeutics","volume":"17","author":"Thakuri","year":"2019","journal-title":"Assay Drug Dev. Technol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1002\/bit.26845","article-title":"3D tumor spheroids as in vitro models to mimic in vivo human solid tumors resistance to therapeutic drugs","volume":"116","author":"Nunes","year":"2019","journal-title":"Biotechnol. Bioeng."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1007\/s00428-017-2289-6","article-title":"Tissue microarray is suitable for scientific biomarkers studies in endometrial cancer","volume":"472","author":"Visser","year":"2018","journal-title":"Virchows Arch."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"335","DOI":"10.1038\/labinvest.2016.151","article-title":"Larger core size has superior technical and analytical accuracy in bladder tissue microarray","volume":"97","author":"Eskaros","year":"2017","journal-title":"Lab. Investig."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1007\/s00428-020-02751-6","article-title":"PD-1\/PD-L1 expression and tumor-infiltrating lymphocytes are prognostically favorable in advanced high-grade serous ovarian carcinoma","volume":"477","author":"Arildsen","year":"2020","journal-title":"Virchows Arch."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1189\/jlb.0109040","article-title":"CD4 and CD8: An inside-out coreceptor model for innate immune cells","volume":"86","author":"Gibbings","year":"2009","journal-title":"J. Leukoc. Biol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1038\/s41416-020-0875-x","article-title":"The untapped potential of ascites in ovarian cancer research and treatment","volume":"123","author":"Ford","year":"2020","journal-title":"Br. J. Cancer"},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"El Harane, S., Zidi, B., El Harane, N., Krause, K.H., Matthes, T., and Preynat-Seauve, O. (2023). Cancer Spheroids and Organoids as Novel Tools for Research and Therapy: State of the Art and Challenges to Guide Precision Medicine. Cells, 12.","DOI":"10.3390\/cells12071001"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"945","DOI":"10.1093\/annonc\/mdl499","article-title":"Pattern and prognostic factors in patients with malignant ascites: A retrospective study","volume":"8","author":"Ayantunde","year":"2007","journal-title":"Ann. Oncol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Capellero, S., Erriquez, J., Battistini, C., Porporato, R., Scotto, G., Borella, F., Di Renzo, M.F., Valabrega, G., and Olivero, M. (2022). Ovarian Cancer Cells in Ascites form Aggregates That Display a Hybrid Epithelial-Mesenchymal Phenotype and Allows Survival and Proliferation of Metastasizing Cells. Int. J. Mol. Sci., 23.","DOI":"10.3390\/ijms23020833"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"2085","DOI":"10.1002\/cam4.4592","article-title":"Clinical significance of CD8-positive lymphocytes on tumor cell clusters of ascites cell block in ovarian high-grade serous carcinoma","volume":"11","author":"Iwahashi","year":"2022","journal-title":"Cancer Med."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Miceska, S., \u0160kof, E., Bu\u010dek, S., Gra\u0161i\u010d Kuhar, C., Ga\u0161ljevi\u0107, G., Smrkolj, \u0160., and Kloboves Prevodnik, V. (2023). The prognostic significance of tumor-immune microenvironment in ascites of patients with high-grade serous carcinoma. Radiol. Oncol., in press.","DOI":"10.2478\/raon-2023-0046"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"e003973","DOI":"10.1136\/jitc-2021-003973","article-title":"Tumor-associated macrophages induced spheroid formation by CCL18-ZEB1-M-CSF feedback loop to promote transcoelomic metastasis of ovarian cancer","volume":"9","author":"Long","year":"2021","journal-title":"J. Immunother. Cancer"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1136\/ijgc-2018-000071","article-title":"Trials in progress: IMagyn050\/GOG 3015\/ENGOT-OV39. A Phase III, multicenter, randomized study of atezolizumab versus placebo administered in combination with paclitaxel, carboplatin, and bevacizumab to patients with newly-diagnosed stage III or stage IV ovarian, fallopian tube, or primary peritoneal cancer","volume":"29","author":"Moore","year":"2019","journal-title":"Int. J. Gynecol. Cancer"},{"key":"ref_30","first-page":"644","article-title":"The oncogenic gene fusion TMPRSS2: ERG is not a diagnostic or prognostic marker for ovarian cancer","volume":"4","author":"Huang","year":"2011","journal-title":"Int. J. Clin. Exp. Pathol."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1530\/ERC-14-0586","article-title":"Expression of ERG protein in prostate cancer: Variability and biological correlates","volume":"22","author":"Ayala","year":"2015","journal-title":"Endocr. Relat. Cancer"}],"container-title":["Cells"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/19\/2390\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:02:53Z","timestamp":1760130173000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2073-4409\/12\/19\/2390"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,9,30]]},"references-count":31,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2023,10]]}},"alternative-id":["cells12192390"],"URL":"https:\/\/doi.org\/10.3390\/cells12192390","relation":{},"ISSN":["2073-4409"],"issn-type":[{"value":"2073-4409","type":"electronic"}],"subject":[],"published":{"date-parts":[[2023,9,30]]}}}