{"doi":"10.1093/biolre/ioaf255","title":"Establishment and characterization of a new immortalized human adenomyosis epithelial-like cell line, tAEC21","abstract":"Adenomyosis occurs when endometrial glands and stroma grow within the uterine myometrium. As a clinically significant disease, adenomyosis causes substantial pelvic pain and heavy menstrual bleeding. It remains understudied due to a lack of translational research tools and model systems. This study aimed to develop a telomerase-transformed, epithelial-like cell line derived from the eutopic endometrium of a subject with focal adenomyosis. De-identified endometrial tissue was processed through mechanical and enzymatic digestion. Epithelial and stromal populations were separated by selective adhesion, followed by fluorescence-activated cell sorting using an epithelial cellular adhesion molecule (EpCAM). EpCAM+ cells were effectively immortalized with the human telomerase reverse transcriptase (TERT) gene. Analyses confirmed the cells were human, free of mycoplasma contamination, and exhibited a unique 16-marker short tandem repeat (STR) profile. Cytogenetic analysis of G-banded metaphase spreads revealed polyploidy with multiple chromosomal rearrangements. The cell line, designated tAEC21, expressed epithelial markers cytokeratin-5 and N-cadherin but not the stromal marker CD10. Cells responded strongly to tumor necrosis factor-alpha stimulation by upregulating interleukin-6, C-X-C motif chemokine ligand 8, C-C motif chemokine ligand 2, and mucin 1 gene expression. In a heterotypic, three-dimensional (3D) spheroid model, tAEC21 formed a biologically relevant structure by creating an epithelial shell around the stromal cell core. In both two-dimensional (2D) monolayer and 3D culture, tAEC21 cells responded to 17β-estradiol (E2) but did not respond to progesterone (P4), consistent with the expression of estrogen (ESR1) and progesterone (PGR) receptors. This new epithelial-like, adenomyosis-derived cell line, tAEC21, will be an impactful, biologically plausible research resource.","journal":"Biology of Reproduction","year":2025,"id":548716,"datarank":0.0,"base_score":0.0,"endowment":0.0,"self_citation_contribution":0.0,"citation_network_contribution":0.0,"self_endowment_contribution":0.0,"citer_contribution":0.0,"corpus_percentile":null,"corpus_rank":null,"citation_count":1,"citer_count":0,"citers_with_citation_signal":0,"citers_with_endowment":0,"datacite_reuse_total":0,"is_dataset":false,"is_dataset_confidence":0.9286,"is_data_producer":false,"deposit_databanks":null,"is_oa":true,"file_count":0,"downloads":0,"has_version_chain":false,"published_date":"2025-01-01","fair_score":null,"fair_percentile":null,"algorithm_id":"datarank_citation_only_1hop_v6","ranking_scope":"data_only","authors":[{"id":1442420,"name":"John Kersey","orcid":"0000-0002-8664-1794","position":1,"is_corresponding":false},{"id":1442421,"name":"Hunter D. Quigley","orcid":"0000-0003-2518-390X","position":2,"is_corresponding":false},{"id":339843,"name":"Shannon M. Hawkins","orcid":"0000-0002-0727-3971","position":3,"is_corresponding":false},{"id":439462,"name":"Yuliya Klymenko","orcid":"0000-0001-7324-8427","position":0,"is_corresponding":true}],"reference_count":116,"raw_metadata":null,"created_at":"2026-07-19T02:54:03.053965Z","pmid":"41263501","pmcid":null,"fwci":null,"citation_percentile":null,"influential_citations":0,"oa_status":null,"license":null,"views":0,"total_file_size_bytes":0,"version_count":0,"fair_f":null,"fair_a":null,"fair_i":null,"fair_r":null,"fair_zscore":null,"fair_rationale":null,"fair_model":null,"fair_agent_version":null,"fair_fulltext_source":null,"fair_has_llm":null,"fair_computed_at":null,"clinical_trials":[],"software_tools":[],"db_accessions":[],"linked_datasets":[],"topics":[]}