Establishment of pathology-validated immortalized patient-derived ovarian endometriosis epithelial cell line

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Researchers established a pathology-validated immortalized ovarian endometriosis epithelial cell line that preserved disease-specific biological features, contrasting it with a teratoma-derived line to highlight the necessity of histopathological classification for rigorous experimental modeling.

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This study established an immortalized patient-derived epithelial cell line from a histopathologically confirmed ovarian endometrioma to evaluate the preservation of disease-specific biological features following lentiviral immortalization. The resulting iEn1 line maintained epithelial characteristics and karyotype stability while demonstrating epithelial-mesenchymal plasticity in xenograft models, though it lacked estrogen receptor beta and interleukin-6 expression. In contrast, a control line derived from a mature cystic teratoma showed chromosomal instability, underscoring that accurate interpretation of cellular models requires integration with definitive histopathological classification. This paper is centrally about endometriosis — specifically the development and validation of a pathology-confirmed immortalized cell line model for ovarian endometriosis.

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Abstract

Endometriosis is a heterogeneous gynecological disorder for which experimental model development is often limited by diagnostic ambiguity and the restricted lifespan of primary epithelial cultures. Although immortalization strategies enable long-term expansion of patient-derived cells, concerns remain regarding the preservation of disease-specific biological characteristics and the risk of model misclassification in the absence of definitive pathological confirmation. In the present study, immortalized ovarian epithelial cell lines from clinically suspected ovarian cystic lesions representing endometriosis were established and it was evaluated whether disease-intrinsic biological features were preserved following immortalization, as determined by definitive histopathological stratification. Primary epithelial cells were isolated from two ovarian cystic lesions and immortalized via lentiviral co-expression of constitutively active cyclin-dependent kinase 4 (CDK4R24C), cyclin D1 and human telomerase reverse transcriptase. One lesion was subsequently identified as an ovarian endometrioma, and the other as a mature cystic teratoma, in postoperative histopathology. The endometriosis-derived immortalized epithelial line iEn1 exhibited sustained proliferative capacity (Cell Counting Kit-8 assay), maintained epithelial lineage characteristics (western blotting for pan-cytokeratin, β-catenin and claudin-1), and retained a stable 46,XX karyotype. These cells demonstrated epithelial-mesenchymal plasticity evidenced by co-expression of epithelial markers with vimentin and N-cadherin, and formed microscopic glandular structures resembling endometriosis-like lesions in an estrogen-supplemented xenograft model. However, immunohistochemistry revealed that estrogen receptor β and interleukin-6, both frequently implicated in endometriosis-associated inflammation, were not detected in either the original tissue or the xenografted lesions, indicating that this particular inflammatory phenotype is not intrinsic to this model. By contrast, the teratoma-derived line iEn2 displayed marked chromosomal instability and phenotypic heterogeneity, demonstrated by karyotyping, consistent with its germ cell origin. Thus, immortalization alone does not determine the biological validity of patient-derived cellular models; rather, accurate interpretation requires integration with definitive histopathological classification. The present study established a pathology-validated immortalized epithelial model of ovarian endometriosis and highlighted the importance of pathology-integrated frameworks for improving experimental rigor in disease modeling.
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Print ISSN: 2049-9434 Online ISSN: 2049-9442 International Journal of Molecular Medicine is an international journal devoted to molecular mechanisms of human disease. International Journal of Oncology is an international journal devoted to oncology research and cancer treatment. Covers molecular medicine topics such as pharmacology, pathology, genetics, neuroscience, infectious diseases, molecular cardiology, and molecular surgery. Oncology Reports is an international journal devoted to fundamental and applied research in Oncology. Experimental and Therapeutic Medicine is an international journal devoted to laboratory and clinical medicine. Oncology Letters is an international journal devoted to Experimental and Clinical Oncology. Explores a wide range of biological and medical fields, including pharmacology, genetics, microbiology, neuroscience, and molecular cardiology. International journal addressing all aspects of oncology research, from tumorigenesis and oncogenes to chemotherapy and metastasis. Multidisciplinary open-access journal spanning biochemistry, genetics, neuroscience, environmental health, and synthetic biology. Open-access journal combining biochemistry, pharmacology, immunology, and genetics to advance health through functional nutrition. Publishes open-access research on using epigenetics to advance understanding and treatment of human disease. An International Open Access Journal Devoted to General Medicine. Article Open Access - Authors: - Article Number: 133|Published online on: September 23, 2026https://doi.org/10.3892/br.2026.2206 - Expand metrics + Endometriosis is a heterogeneous gynecological disorder for which experimental model development is often limited by diagnostic ambiguity and the restricted lifespan of primary epithelial cultures. Although immortalization strategies enable long‑term expansion of patient‑derived cells, concerns remain regarding the preservation of disease‑specific biological characteristics and the risk of model misclassification in the absence of definitive pathological confirmation. In the present study, immortalized ovarian epithelial cell lines from clinically suspected ovarian cystic lesions representing endometriosis were established and it was evaluated whether disease‑intrinsic biological features were preserved following immortalization, as determined by definitive histopathological stratification. Primary epithelial cells were isolated from two ovarian cystic lesions and immortalized via lentiviral co‑expression of constitutively active cyclin‑dependent kinase 4 (CDK4R24C), cyclin D1 and human telomerase reverse transcriptase. One lesion was subsequently identified as an ovarian endometrioma, and the other as a mature cystic teratoma, in postoperative histopathology. The endometriosis‑derived immortalized epithelial line iEn1 exhibited sustained proliferative capacity (Cell Counting Kit‑8 assay), maintained epithelial lineage characteristics (western blotting for pan‑cytokeratin, β‑catenin and claudin‑1), and retained a stable 46,XX karyotype. These cells demonstrated epithelial‑mesenchymal plasticity evidenced by co‑expression of epithelial markers with vimentin and N‑cadherin, and formed microscopic glandular structures resembling endometriosis‑like lesions in an estrogen‑supplemented xenograft model. However, immunohistochemistry revealed that estrogen receptor β and interleukin‑6, both frequently implicated in endometriosis‑associated inflammation, were not detected in either the original tissue or the xenografted lesions, indicating that this particular inflammatory phenotype is not intrinsic to this model. By contrast, the teratoma‑derived line iEn2 displayed marked chromosomal instability and phenotypic heterogeneity, demonstrated by karyotyping, consistent with its germ cell origin. Thus, immortalization alone does not determine the biological validity of patient‑derived cellular models; rather, accurate interpretation requires integration with definitive histopathological classification. The present study established a pathology‑validated immortalized epithelial model of ovarian endometriosis and highlighted the importance of pathology‑integrated frameworks for improving experimental rigor in disease modeling. Copy and paste a formatted citation Spandidos Publications style Saisomboon S, Srikummoon N, Pattanachot J, Kleebkaow P, Amnatbuddee S, Deenonpoe R, Wichian P, Sridakhun N, Eakpinitpitaya T, Chumworathayi B, Chumworathayi B, et al: Establishment of pathology‑validated immortalized patient‑derived ovarian endometriosis epithelial cell line. Biomed Rep 25: 133, 2026. APA Saisomboon, S., Srikummoon, N., Pattanachot, J., Kleebkaow, P., Amnatbuddee, S., Deenonpoe, R. ... Vaeteewoottacharn, K. (2026). Establishment of pathology‑validated immortalized patient‑derived ovarian endometriosis epithelial cell line. Biomedical Reports, 25, 133. https://doi.org/10.3892/br.2026.2206 MLA Saisomboon, S., Srikummoon, N., Pattanachot, J., Kleebkaow, P., Amnatbuddee, S., Deenonpoe, R., Wichian, P., Sridakhun, N., Eakpinitpitaya, T., Chumworathayi, B., Seubwai, W., Thanan, R., Pinlaor, S., Okada, S., Vaeteewoottacharn, K."Establishment of pathology‑validated immortalized patient‑derived ovarian endometriosis epithelial cell line". Biomedical Reports 25.5 (2026): 133. Chicago Saisomboon, S., Srikummoon, N., Pattanachot, J., Kleebkaow, P., Amnatbuddee, S., Deenonpoe, R., Wichian, P., Sridakhun, N., Eakpinitpitaya, T., Chumworathayi, B., Seubwai, W., Thanan, R., Pinlaor, S., Okada, S., Vaeteewoottacharn, K."Establishment of pathology‑validated immortalized patient‑derived ovarian endometriosis epithelial cell line". Biomedical Reports 25, no. 5 (2026): 133. https://doi.org/10.3892/br.2026.2206

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