Novel 3D in vitro models to evaluate trophoblast migration and invasion

Abstract

ProblemEmbryo implantation depends on the interactions between the developing embryo and the maternal endometrium. Signals originating from the decidua play a critical role in the process of implantation and trophoblast invasion; however, the molecular mechanisms mediating this interaction are poorly understood. The objective of this study was to develop in vitro models that would mimic the processes of attachment, migration, and early invasion of the trophoblast.Methods of studyFirst trimester trophoblast cells (Sw.71 cells) were cultured in low attachment plates to form blastocyst‐like spheroids (BLS). Epithelial‐mesenchymal transition (EMT) characterization during BLS formation was determined by RT‐PCR and Western Blot. The two 3D in vitro culture models consist of (a) trophoblast migration: BLS cultured in suspension (b) trophoblast invasion: human endometrium stromal cells (HESC) plated in the bottom of a 96‐well plate, covered by Matrigel and BLS transferred on top. Matrigel was used to mimic the human endometrial extracellular matrix.ResultsUsing 3D cell culture systems and real‐time imaging, we are able to determine the impact of endometrial factors on trophoblast cell function. Endometrial stromal cells promote blastocyst‐like spheroid migration of trophoblast cells and invasion of the extracellular matrix.ConclusionWe report the characterization of 3D in vitro models to evaluate the interaction between endometrial cells and trophoblast during the process of migration and invasion. The models are useful tools in order to further study the molecular mechanism of embryo‐maternal uterine cells interactions.

Differential Secretion of Inflammatory Cytokines by Human Trophoblasts in the Presence of <b><i>Escherichia coli</i></b>, <b><i>Lactobacillus crispatus</i></b>, and <b><i>Lactobacillus jensenii</i></b>

“…The differential fold increase in cytokine/chemokine levels in the presence of bacteria was less likely to be explained by normal variations (Table 1). Sw.71 is a well-characterized cell lineage that has been accepted as a study model for human trophoblast biology [54,57]. Studies have shown that Sw.71 secretes an array of cytokines when stimulated with different bacterial products [58,59].…”

Modelling the Human Placental Interface In Vitro—A Review

“…Recently, the inclusion of self-assembled spheroids of extravillous trophoblasts has allowed for complex 3D cell-cell interactions, bringing important insight into the mechanisms underlying trophoblast migration and invasion (Figure 3c). For instance, You et al demonstrated that endometrial signaling is essential to promote and guide trophoblast invasion by observing that trophoblasts were able to migrate from the spheroids and invade the Matrigel only when there was an underlying layer of human endometrial stromal cells [36]. bringing important insight into the mechanisms underlying trophoblast migration and invasion (Figure 3c).…”

Detrimental effects of lipopolysaccharide on the attachment and outgrowth of various trophoblastic spheroids on human endometrial epithelial cells

“…Many in vitro models have provided insights into the implantation process [19,20]. In a recent study, in vitro implantation models were designed using the spheroid form of trophoblastic cells to surrogate embryos [21].…”