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¹ Department of Human Anatomy and Cell Science, University of Manitoba, Winnipeg, Manitoba, Canada
² Department of Immunology, Martin Luther University, Halle-Wittenberg, Germany
³ Department of Obstetrics and Gynecology, University of Aberdeen, Aberdeen, UK
⁴ Department of Anatomy, RWTH Aachen, Germany
⁵ Department of Dermatology, RWTH Aachen, Germany
⁶ Department of Analytical and Environmental Chemistry, Martin Luther University, Halle-Wittenberg, Germany
⁷ Department of Pathology, Martin Luther University, Halle-Wittenberg, Germany

(Requests for offprints should be addressed to T Klonisch, Department of Human Anatomy and Cell Science, Faculty of Medicine, 130 Basic Medical Sciences, 730 Williams Avenue, University of Manitoba, Winnipeg, Manitoba, R3E 0W3, Canada; Email: klonisch{at}cc.umanitoba.ca)

Information on the regulation of steroid hormone receptors and their distinct functions within the human endometrial epithelium is largely unavailable. We have immortalized human primary endometrial epithelial cells (EECs) isolated from a normal proliferative phase endometrium by stably transfecting the catalytic subunit (hTERT) of the human telomerase complex and cultured these hTERT-EECs now for over 350 population doublings. Active hTERT was detected in hTERT-EECs employing the telomerase repeat amplification assay protocol. hTERT-EECs revealed a polarized, non-invasive epithelial phenotype with apical microvilli and production of a basal lamina when grown on a three-dimensional collagen–fibroblast lattice. Employing atomic force microscopy, living hTERT-EECs were shown to produce extracellular matrix (ECM) components and ECM secretion was modified by estrogen and progesterone (P4). hTERT-EECs expressed inducible and functional endogenous estrogen receptor-alpha (ER-alpha) as demonstrated by estrogen response element reporter assays and induction of P4 receptor (PR). P4 treatment down-regulated PR expression, induced MUC-1 gene activity and resulted in increased ER-beta transcriptional activity. Gene activities of cytokines and their receptors interleukin (IL)-6, leukemia inhibitory factor (LIF), IL-11 and IL-6 receptor (IL6-R), LIF receptor and gp130 relevant to implantation revealed a 17 beta-estradiol (E2)-mediated up-regulation of IL-6 and an E2- and P4-mediated up-regulation of IL6-R in hTERT-EECs. Thus, hTERT-EECs may be regarded as a novel in vitro model to investigate the role of human EECs in steroid hormone-dependent normal physiology and pathologies, including implantation failure, endometriosis and endometrial cancer.

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