Androgens Modulate the Morphological Characteristics of Human Endometrial Stromal Cells Decidualized In Vitro

Abstract

The activated androgen receptor (AR) in decidualizing human endometrial stromal cells (HESCs) regulates genes involved in cytoskeletal organization, cell motility, and cell cycle progression. Androgens also enhance the secretion of prolactin, a widely used marker of decidualized HESCs. The purpose of the present study was to investigate the direct effects of androgens on the ultrastructural changes associated with decidual transformation of HESCs. Primary HESC cultures were established and propagated, and confluent cultures were decidualized for 6 days with 8-bromoadenosine 3',5'-cyclic monophosphate (8-br-cAMP) and progesterone (P4) in the presence or absence of dihydrotestosterone (DHT). Phase-contrast image analysis demonstrated that DHT increases the shape index of decidualizing cells, which was reversed upon cotreatment with the AR antagonist flutamide. Electron microscopy demonstrated that DHT enhances many of the ultrastructural changes induced by 8-br-cAMP and P4 in HESCs. Decidualizing cells are characterized by an abundant cytoplasm, multiple cell surface projections and, unlike undifferentiated HESCs, form 2 or more cell layers. The DHT further stimulated cytoplasmic expansion, lipid droplet formation, the production of an abundant extracellular matrix, and gap junction formation in decidualized HESCs. The present study demonstrates that androgen signaling has an impact on the morphological and ultrastructural changes associated with the decidual process. Our findings show that androgens promote the development and expansion of cytoplasmic organelles and gap junctions in decidualizing HESCs. These results suggest that androgens in early pregnancy play an important role in promoting the cellular transformation associated with decidualization.

Loss of miR-542-3p enhances IGFBP-1 expression in decidualizing human endometrial stromal cells

“…The efficiency of decidualization in primary HESC cultures was confirmed by observing morphological changes and by measuring PRL secretion, the principal decidual marker1011. HESCs exhibited a spindle-shaped fibroblast-like appearance when propagated in culture but acquired the typical morphology of decidual cells, i.e.…”

Exposure of decidualized HIESC to low oxygen tension and leucine deprivation results in increased IGFBP-1 phosphorylation and reduced IGF-I bioactivity

“…The morphological characteristic of HIESC was determined before and after decidualization using the approach of calculating the shape index as described (Kajihara et al, 2014). Phase contrast images were captured 6 days after decidualization (treatment with cAMP + MPA).…”

“…Individual cells (50 cells) were outlined from three cultures (n = 3) for control and treated cells and the roundness factor was determined using Image Pro Premier software. The shape index formula used was as reported by Kajihara et al (2014) (4pi x area/perimeter 2 ). A circular object would have a shape index of 1 whereas a straight line has an index of 0.…”

Bisphenol A impairs decidualization of human uterine stromal fibroblasts

“…In humans, this process is under maternal control, meaning that decidualization occurs every cycle in the mid- to late-secretory phase due to endocrine and paracrine regulation. Decidual cells are characterized by rounded nuclei, a large cytoplasmic area, significant extracellular matrix production and gap junction formations [17]. Prolactin and IGFBP-1 are produced and secreted by decidual cells and are considered markers to assess the status of stromal cells decidualizing in vitro [16].…”