Directed biomechanical compressive forces enhance fusion efficiency in model placental trophoblast cultures.

The syncytiotrophoblast is a multinucleated structure that arises from fusion of mononucleated cytotrophoblasts, to sheath the placental villi and regulate transport across the maternal-fetal interface. Here, we ask whether the dynamic mechanical forces that must arise during villous development might influence fusion, and explore this question using in vitro choriocarcinoma trophoblast models. We demonstrate that mechanical stress patterns arise around sites of localized fusion in cell monolayers, in patterns that match computational predictions of villous morphogenesis. We then externally apply these mechanical stress patterns to cell monolayers and demonstrate that equibiaxial compressive stresses (but not uniaxial or equibiaxial tensile stresses) enhance expression of the syndecan-1 and loss of E-cadherin as markers of fusion. These findings suggest that the mechanical stresses that contribute towards sculpting the placental villi may also impact fusion in the developing tissue. We then extend this concept towards 3D cultures and demonstrate that fusion can be enhanced by applying low isometric compressive stresses to spheroid models, even in the absence of an inducing agent. These results indicate that mechanical stimulation is a potent activator of cellular fusion, suggesting novel avenues to improve experimental reproductive modelling, placental tissue engineering, and understanding disorders of pregnancy development.

In Situ Measurement of Intra-tumoral Tissue Rigidity.

Local tissue scale mechanical properties are essential for understanding cell fate and function; however, few methods to measure stiffness at this length scale exist, and applications in 3D tissues can present further challenges. To address this need, microgel-based sensors fabricated out of the thermally responsive hydrogel poly(N-isopropylacrylamide) were developed allowing internal architectures of tissues to be mapped by optically measuring microgel response when actuated in a matrix. These robust probes are widely applicable for in vitro and in vivo studies of tissue mechanics providing tissues can be fluorescently imaged. Here we describe the fabrication of these thermally responsive hydrogel sensors, calibration of the microgels using phantom tissues, and image processing techniques used to make the measurements.