Sex Steroids Regulate Epithelial–Stromal Cell Cross Talk and Trophoblast Attachment Invasion in a Three-Dimensional Human Endometrial Culture System

Abstract

Human embryo implantation involves a complex network of molecular signaling that is modulated by endocrine and paracrine pathways. Here, we performed studies using a unique and recently developed three-dimensional (3D) implantation model, characterized by an endometrium-like 3D culture system and Jar cell-derived spheroids mimicking the embryo/trophoblast. The aims were to investigate the effects of 17β estradiol (E2) and medroxyprogesterone acetate (MPA) on (1) the interaction between epithelial and stromal cells, and (2) the attachment and invasion of trophoblast cells. We observed that epithelial and stromal cells in the 3D culture were ERα⁺, ERβ⁺, and PR⁺. Decidualization was confirmed by enhanced prolactin gene expression on day 7 of E2 plus MPA treatment. An effect of epithelial cells on the decidualization of stromal cells was indicated by significantly higher levels of prolactin mRNA expression in the 3D culture compared to stromal cells grown within the fibrin-agarose gel matrix. On the other hand, the relative gene expressions of E-cadherin and IL-1β in epithelial cells of the 3D culture under decidualization conditions significantly differed from those in epithelial cells grown over the fibrin-agarose gel matrix without stromal cells, pointing to regulation of epithelial cells by the stroma. The attachment rate of Jar spheroids to the 3D was significantly increased by E2 plus MPA treatment. Analyses of Z-stack confocal and stained optic microscopic images demonstrated that Jar spheroids breached the epithelial cell monolayer, invaded, and were embedded into the 3D matrix in response to decidualization signals. In summary, the newly bioengineered system provides a unique model for studying interactions between the different endometrial cell compartments, via soluble-paracrine signals as well as cell-to-cell interactions, and is a useful tool to study early embryonic implantation events.

Expression of Aquaporins in Human Embryos and Potential Role of AQP3 and AQP7 in Preimplantation Mouse Embryo Development

“…JAr spheroids represent an appropriate embryo-like model for studies mimicking implantation [17]. We found that JAr cells expressed AQP3 (Fig.…”

Functional Antagonism between High Temperature Requirement Protein A (HtrA) Family Members Regulates Trophoblast Invasion

“…Ishikawa cells derived from endometrial adenocarcinoma and T-HESCs established from primary HESCs by telomerase immortalization are well accepted for in vitro studies of human endometrium (17)(18)(19)(20). Specifically, both cell lines share similar biochemical properties with primary endometrial cells when induced by ovarian steroids and cAMP.…”

Local remodeling of synthetic extracellular matrix microenvironments by co-cultured endometrial epithelial and stromal cells enables long-term dynamic physiological function

“…Hence, defining an appropriate ECM hydrogel for engineering mucosal barrier tissues for extended in vitro culture and function is an ongoing challenge. In epithelial monocultures, the basement membrane extract Matrigel™ is commonly used, 13,14 whereas type I collagen gels or fibrin gels are used as representative microenvironments for stromal cells, 15–17 with no single composition ideal for both cell types in co-culture. Further, although these and other natural ECM gels are readily available commercially, they exhibit substantial inherent batch-to-batch variability, variations in growth factor composition depending on preparation method, limited opportunity to tailor matrix degradation or signaling properties, and are difficult to microfabricate for organs-on-chips type applications.…”