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Hum. Reprod. Advance Access published online on January 21, 2005
Human Reproduction, doi:10.1093/humrep/deh722
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Human Reproduction © European Society of Human Reproduction and Embryology 2005; all rights reserved
Received August 18, 2004
Accepted December 7, 2004

Article

A novel organotypic culture model for normal human endometrium: regulation of epithelial cell proliferation by estradiol and medroxyprogesterone acetate

M. Bläuer 1*, P.K. Heinonen 2, P.M. Martikainen 3, E. Tomás 2, and T. Ylikomi 1

1 Department of Cell Biology, Medical School, 33014 University of Tampere, Tampere, Finland
2 Department of Obstetrics and Gynecology, Tampere University Hospital, Tampere, Finland
3 Department of Pathology, Centre for Laboratory Medicine, Tampere University Hospital, 33521Tampere, Finland

* To whom correspondence should be addressed.
M. Bläuer, E-mail: Merja.Blauer{at}uta.fi


  Abstract

BACKGROUND: A novel organotypic culture system was established for modelling the hormonal responses of the normal human endometrium in vitro. METHODS: Endometrial epithelial cells were cultured as glandular organoids within reconstituted extracellular matrix (Matrigel) in tissue culture inserts and stromal cells on plastic below the epithelial compartment. The effects of estradiol (E2) and E2 together with medroxyprogesterone acetate (MPA) on cell proliferation and the expression of estrogen receptor {alpha} (ER{alpha}) and progesterone receptor (PR) were studied in 10 epithelial-stromal co-cultures and in three parallell monocultures of epithelial organoids. RESULTS: In co-cultures, E2 was shown to increase the percentage of Ki67-positive cells by ~2-fold relative to untreated controls. In the presence of MPA, a significant decrease in cell proliferation was detected. Similar results were obtained when the corresponding percentages of Ki67-positive organoids were calculated instead of individual cells. In the absence of stromal fibroblasts, Ki67 epithelial labelling remained below the control value after both hormonal treatments. Epithelial organoids retained their capacity to express estrogen and progesterone receptors in culture. E2 was shown to markedly increase and MPA to down-regulate the expression of PR. The expression of ER{alpha} was only slightly affected by either hormonal treatment. CONCLUSIONS: The present organotypic model provides a novel in vitro system in which to study the effects of steroids in the normal human endometrium both in terms of cell proliferation and gene expression. The culture system holds promise as a useful method to screen novel steroid compounds and may help to circumvent problems related to the use of animal models.

Keywords: endometrium; human; Matrigel; organoid; organotypic culture.
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