A 3D approach to reproduction.

For over a century, 2D cell culture has been extensively used for all the different research fields. However, this in vitro system does not allow to reproduce the natural structures of the original tissue, causing several changes and, in most cases, the loss of cell-to-cell communications and cell-to-extracellular matrix interactions. Based on this, during the last years, novel 3D platforms, able to mimic the in vivo milieu, are being developed. The advantages of the use of 3D models are: the reduction of the gap between cell culture and physiological environment; imitation of the specific architecture; partially maintenance of the mechanical and biochemical cues of the original tissue. Currently, 3D systems are used in a broad range of studies, including the field of reproduction, where they have been applied to promote maturation of follicles and oocytes and embryo culture. Here, we review 2D and 3D cell culture methods, discussing advantages and limitations of these techniques. We report the fundamental mechanisms involved in cell ability to perceive and respond to mechanical cues and their role in transmitting signals to and between cells and in regulating intracellular signaling pathways. In particular, we focus on the main effectors of the Hippo pathway, Yes-associated protein (YAP) and WW domain-containing transcription regulator protein 1 (TAZ), describing their behavior and function in oocytes and embryos. Lastly, we provide an overall perspective of the most recent 3D technologies developed in the field of reproduction, describing how their use may revolutionize the understanding of cellular behavior and provide novel tools, useful in reproductive technologies and livestock production.