ALBERT

Description: Biofabrication, including printing technologies, has emerged as a powerful approach to the design of disease models, such as in cancer research. In breast cancer, adipose tissue has been acknowledged as an important part of the tumor microenvironment favoring tumor progression. Therefore, in this study, a 3D-printed breast cancer model for facilitating investigations into cancer cell-adipocyte interaction was developed. First, we focused on the printability of human adipose-derived stromal cell (ASC) spheroids in an extrusion-based bioprinting setup and the adipogenic differentiation within printed spheroids into adipose microtissues. The printing process was optimized in terms of spheroid viability and homogeneous spheroid distribution in a hyaluronic acid-based bioink. Adipogenic differentiation after printing was demonstrated by lipid accumulation, expression of adipogenic marker genes, and an adipogenic ECM profile. Subsequently, a breast cancer cell (MDA-MB-231) compartment was printed onto the adipose tissue constructs. After nine days of co-culture, we observed a cancer cell-induced reduction of the lipid content and a remodeling of the ECM within the adipose tissues, with increased fibronectin, collagen I and collagen VI expression. Together, our data demonstrate that 3D-printed breast cancer-adipose tissue models can recapitulate important aspects of the complex cell–cell and cell–matrix interplay within the tumor-stroma microenvironment.

Description: Important processes of living cells, including intracellular transport, cell crawling, contraction, division, and mechanochemical signal transduction, are controlled by cytoskeletal (CSK) dynamics. CSK dynamics can be measured by tracking the motion of CSK-bound particles. Particle motion has been reported to follow a superdiffusive behavior that is believed to arise from ATP-driven intracellular stress fluctuations generated by polymerization processes and motor proteins. The power spectrum of intracellular stress fluctuations has been suggested to decay with 1/2 (Lau et al, Phys Rev Lett 91:198101). Here we report direct measurements of cellular force fluctuations that are transmitted to the extracellular matrix, and compared them with the spontaneous motion of CSK-bound beads. Fibronectin coated fluorescent beads (Ø 1 m) were bound to the CSK of confluent human vascular endothelial cells. Forces transmitted to the extracellular matrix (ECM) were quantified by plating these cells onto a collagen coated elastic polyacrylamide hydrogel, and measuring the gel deformation from the displacement of embedded fluorescent beads (Ø 0.5 m). Bead motion of both CSK-bound and ECM-bound beads were measured with nanometer-resolution and expressed as mean square displacement (MSD). The MSD of both CSK-bound and ECM-bound beads displayed a superdiffusive behavior that was well described by a power law: MSD = a*t^b. Surprisingly, we found an identical power law exponent for both CSK-bound and ECM-bound beads of b = 1.6. This finding suggests that the spontaneous motion of CSK-bound beads is driven by stress fluctuations with a 1/ b+1 power spectrum. This result is consistent with the notion that CSK dynamics and CSK stress fluctuations are closely coupled.