Establishment and characterization of cell lines from human endometrial epithelial and mesenchymal cells from patients with endometriosis

Abstract Objective To establish and characterize cell lines derived from human endometrial epithelial cells (ECs) and mesenchymal cells (MCs) from patients with and without endometriosis. Design In vitro experimental study Setting University and National Cancer Center Research Institute Patients Two women with endometriosis and two women without endometriosis Intervention(s) Sampling of endometrial ECs and MCs Main Outcome Measure(s) We established immortalized endometrial ECs and MCs. Quantitative reverse transcription-polymerase chain reaction (qRT-PCR), immunocytochemical analysis, and RNA sequence profiling were performed to characterize immortalized cells. A cell proliferation assay, three-dimensional culture, and assays for hormonal responses were performed to characterize the features of ECs. Results qRT-PCR, immunocytochemical analysis, and western blotting revealed that ECs and MCs maintained their original features. Moreover, immortalized cells were found to retain responsiveness to sex steroid hormones. ECs formed a gland-like structure in 3D culture, indicating the maintenance of normal EC phenotypes. RNA sequence profiling, principal component analysis, and clustering analysis showed that the gene expression patterns of immortalized cells were different from those of cancer cells. Several signaling pathways that were significantly enriched in ECs and MCs with endometriosis were revealed. Conclusion We successfully obtained four paired immortalized endometrial ECs and MCs from patients with and without endometriosis. Using these cells could help identify diagnostic and therapeutic targets for endometriosis. The cell lines established in this study will thus serve as powerful experimental tools in the study of endometriosis.