Deciphering embryo-maternal cross-talk: advanced in vitro approaches

Santos, Gislaine dosMillán de la Blanca, María GemmaCajas, Yulia NathalyMazzarella, RosaneCañón-Beltrán, KarinaGonzalez Martínez, Maria EncinaRizos, Dimitrios

Abstract Embryo-maternal communication is a critical process that influences early embryonic development, implantation success, and pregnancy outcomes across mammalian species. This review examines the diverse in vitro systems developed to study this complex dialogue, highlighting their applications, advantages, and limitations. We explore conventional approaches such as two-dimensional (2D) cell cultures, which despite their simplicity, face challenges in replicating the three-dimensional (3D) architecture and cellular functions present in vivo. The review progresses through increasingly sophisticated models, including fluid co-culture systems that incorporate bioactive molecules, explant cultures that maintain tissue architecture, air-liquid interface systems that promote epithelial polarization and differentiation, 3D organoid systems that recapitulate complex structural organization, and organ-on-a-chip platforms that recreate mechanical forces and dynamic conditions. Special attention is given to the emerging role of extracellular vesicles (EVs) as mediators of embryo-maternal communication, transporting crucial molecular signals between the embryo and reproductive tract. By comparing these systems across species and developmental stages, we provide a comprehensive framework for selecting appropriate models based on specific research questions. The integration of these in vitro approaches with advanced analytical techniques offers promising avenues for understanding embryo-maternal cross-talk, potentially leading to improved assisted reproductive technologies and strategies to mitigate early pregnancy loss. As technology advances, the continued refinement of these systems will further illuminate the intricate molecular and cellular mechanisms underlying successful embryo development and implantation.