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.
Highlights
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
We implemented two experimental methods for our cell motility experiments with HSH substrates: horizontal HSH experiments, where cells moved across the width of the strip regions, and vertical HSH experiments, where cells moved along the length of the strip regions (Fig. 1A,B)
We first investigated the motility of A549 epithelial cells and WI-38 fibroblasts attached on uniform substrates with different stiffnesses over time
Summary
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. Examining how cells interact with each other in response to mechanically differential substrates would allow an increased understanding of their coordinated cell motility. We cultured WI-38 fibroblasts and A549 epithelial cells to probe their motile response to the substrates. 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. In the co-culture system, the WI-38 fibroblasts moved more toward the soft area of the HSH substrate when compared to A549 epithelial cells, indicating the HSH system may be useful for separation of different cell types. Our approach indicates that in addition to cell separation, the HSH substrate may be able tto spatially and mechanically guide cells for controlled motility in co-culture
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