Effects of steroid hormones on differentiated glandular epithelial and stromal cells in a three dimensional cell culture model of the canine endometrium

Abstract

BackgroundOestrogens and progesterone have a significant impact on the endometrium during the canine oestrous cycle. Their receptors mediate plasma steroid hormone levels and are expressed in several endometrial cell types. Altered steroid receptor expression patterns are involved in serious uterine diseases; however the mechanisms of hormone action during pathogenesis in these tissues remain unclear. The development of 3D culture systems of canine endometrial cells provides an opportunity for the effects of steroid hormones to be quantitatively assessed in a more in vivo-like setting. The present study aimed to determine the effects of the steroid hormones 17β-estradiol (E) and progesterone (P) on the expression of the oestrogen and progesterone receptors (ER and PR), and on proliferative activity, in a 3D co-culture system of canine uterine origin, comprising differentiated endometrial glands, and stromal cells (SCs).ResultsMorphology, differentiation, and apical-basolateral polarity of cultured glandular epithelial cells (GECs) were comparable to those in native uterine tissue as assessed by immunohistochemistry using differentiation markers (β-catenin, laminin), lectin histochemistry, and transmission electron microscopy. Supplementation of our 3D-culture system with E (at 15, 30 and 100 pg/mL) resulted in constant levels of ER expression in GECs, but reduced expression levels in SCs. PR expression was reduced in both GECs and SCs following treatment with E. 3 ng/mL P resulted in increased ER expression in GECs, but a decrease in SCs. PR expression in GECs increased in all P-treated groups, whereas PRs in SCs decreased with the lowest and highest doses, but increased with the middle dose of treatment. Proliferative activity, assessed by Ki67 staining, remained below 1% in all assays and cell types.ConclusionsThe present study demonstrates the applicability of our 3D organotypic canine endometrium-derived culture system for cellular-level studies. 3D cultures represent near-physiological systems allowing reproducible quantitative experimentation, thus reducing the need to experiment on living animals. The results of the present investigation emphasize the importance of co-culture of the uterine glands with SCs, as it was shown that the responsiveness of the different cell types to steroid hormones were divergent in the 3D cell culture model.

Köpeklerde Endotoksik Pyometranın Klinik, Bakteriyolojik ve Histopatolojik Özellikleri

“…In addition, suppression in bone marrow activity which leads to anemia and remarkably increased leukocytes can occur due to the toxin effect in bitches with pyometra related to endotoxemia [11,12] . Besides, the prognosis of pyometra may vary according to the absorption of bacterial toxin and the histomorphological alterations of the uterus [2,[13][14][15][16] . Therefore, the determination of the blood endotoxin level and histopathological changes in the uterus are more important for estimating the prognosis of the disease in bitches with pyometra [2,14,17] .…”

Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures

“…Spheroids are appropriate units for tissue reconstruction. Often, they are developed using 3D scaffolds consisting of porous materials or from extracellular matrix (ECM) proteins (Tan et al., ; Bartel et al., ; Lawrenson et al., ; Matsusaki et al., ). However, the application of scaffold materials alters cell morphology and physiology.…”

Steroid Hormonların Yumurtacı Tavukların Epitel ve Stroma Hücre Kültürüne İn vitro Etkileri

“…Thus, both the endometrial epithelial and stromal cells culture are important methods that can be used to explore the physiological function of the uterus of the poultry. Currently, previous studies have successfully isolated and cultured the endometrial epithelial and stromal cells in most mammals, such as cattle [23] , canine [24] , equine [25] and rabbit [26] and so on. It is also mentionable that Park et al [27] have isolated and cultured human endometrial epithelial and stromal cells obtained by using a centrifugation method [27] , and interestingly enough, Bläuer et al [28] successfully constructed a novel organotypic culture model for normal human endometrium by co-culture in vitro [28] .…”