Probing coordinated co-culture cancer related motility through differential micro-compartmentalized elastic substrates

Abstract

Cell development and behavior are driven by internal genetic programming, but the external microenvironment is increasingly recognized as a significant factor in cell differentiation, migration, and in the case of cancer, metastatic progression. Yet it remains unclear how the microenvironment influences cell processes, especially when examining cell motility. One factor that affects cell motility is cell mechanics, which is known to be related to substrate stiffness. Examining how cells interact with each other in response to mechanically differential substrates would allow an increased understanding of their coordinated cell motility. In order to probe the effect of substrate stiffness on tumor related cells in greater detail, we created hard–soft–hard (HSH) polydimethylsiloxane (PDMS) substrates with alternating regions of different stiffness (200 and 800 kPa). We then cultured WI-38 fibroblasts and A549 epithelial cells to probe their motile response to the substrates. We found that when the 2 cell types were exposed simultaneously to the same substrate, fibroblasts moved at an increased speed over epithelial cells. Furthermore, the HSH substrate allowed us to physically guide and separate the different cell types based on their relative motile speed. We believe that this method and results will be important in a diversity of areas including mechanical microenvironment, cell motility, and cancer biology.

Cell Migration Regulated by Spatially Controlled Stiffness inside Composition‐Tunable Three‐Dimensional Dextran Hydrogels

“…From the migration distance measured in this study, cells moved at the highest speed in S2 gels with uniform stiffness (Figure 9a), consistent with a previous finding. [53] It has been demonstrated that cell migration through the ECM is accompanied by changes in ECM structure. [54,55] As the stiffness of gels increase, more cellular work is required to cause matrix displacement, which entails higher energetic costs.…”

Development of an Optical System for Strain Drop Measurement of Osteosarcoma Cells on Substrates with Different Stiffness

“…Subsequently, with the aim to perform a validation of the sensitivity of the measurement technique, we measured the strain drop of untreated 143B cells in comparison to 143B cells previously treated with glutaraldehyde (GA) (3% in PBS) [ 15 , 48 , 49 ] to make the cells stiffer than the control cells by creating chemical bonds between the cell and the substrate [ 50 ], or with cytochalasin-D (CD) (0.5 μg/mL in DMSO) to inhibit the assembly of actin filaments, reducing the cells’ ability to withstand stress [ 51 ] when plated on 5 kPa substrates. GA and CD treatments lasted 20 min and 15 min before the beginning of experimental testing, respectively.…”

Discrimination between NSIP- and IPF-Derived Fibroblasts Based on Multi-Parameter Characterization of Their Growth, Morphology and Physic-Chemical Properties

“…It should be noted here that PDMS surface is highly hydrophobic [79], thus the ability of cells to adhere to this substrate is usually reduced. To overcome this problem, the PDMS substrates are usually modified, e.g., by coating with ECM proteins [80][81][82][83]. However, the spreading of cells on unmodified PDMS is possible for some cells [75,84], most likely due to the presence of serum proteins in the culture medium, serving as an adhesion matrix.…”