Exploiting Substrate Cues for Co-Culturing Cells in a Micropattern

Abstract

Spatial distribution of cells and their interactions between neighboring cells in native microenvironments are of fundamental importance in determining cell fate decisions such as migration, growth, and differentiation. Controlling the spatial distribution of different cell types in defined geometries can replicate these native environments, which can be a useful model for several studies. While spatiotemporal control over multiple cell arrangements is required to achieve the complex tissue architecture, unfortunately, conventional cell patterning techniques usually allow only single patterning with a single cell type. In the present study, we introduce a simple lithographic method to pattern multiple cell types in a spatially controlled manner by utilizing the biophysical cues present at the corners of the patterned geometry. By fabricating micropatterns of different shapes, we demonstrate how the cell can be constrained to pattern along the corners of patterned geometries owing to the presence of topographical cues, leaving empty voids in the center that can be further utilized for patterning a second cell type. We also demonstrate that the cell alignment along the pattern is a dynamic process and the cells migrate from a more uniform cell-adhesive region toward the topographical cues. The cytoskeleton arrangement was geometry-dependent, which was confirmed through a series of in vitro evaluations, such as scanning electron microscopy and fluorescence microscopy. These findings have not only helped us in exploring the importance of these cues in guiding the cell fate but have also allowed us to develop a technique, which self-patterns the cells without any expensive exogenous cues and can be used as a model protocol to eventually organize cells into a specific pattern with micron-scale precision in vitro.

Exploiting the Biophysical Cues toward Dual Differentiation of hMSC’s within Geometrical Patterns

“…In our previous study, we have demonstrated the effect of these geometrical cues toward the behavior of osteoblast and fibroblast cells and utilized these effects to develop a model to coculture cells, but the effect of these cues in guiding a biological process such as differentiation was not studied. 32 The study also demonstrated how these cues can influence cellular morphology between the cells present at the center vs the edges and how these patterns can be transmitted to a cell seeded after a time interval, which further allows self-patterning of a second cell type. We hypothesized that cell alignment at the corner/edges signals the subsequently seeded cells to align according to the pattern due to cell−cell signaling.…”

3D Bioprinted Alginate-Silk-Based Smart Cell-Instructive Scaffolds for Dual Differentiation of Human Mesenchymal Stem Cells

“…One of the main characteristics of a biomaterial is the ability to support cell adhesion, proliferation, and migration without causing any cytotoxic effects. 49 The viability of the cells were studied via live (calcein AM) and dead (propidium iodide, PI) staining. As seen from Figure 3a, the cells showed increased viability in all three scaffolds (AL, AS, ApS) after 7 days of culture.…”

“…One of the main characteristics of a biomaterial is the ability to support cell adhesion, proliferation, and migration without causing any cytotoxic effects . The viability of the cells were studied via live (calcein AM) and dead (propidium iodide, PI) staining.…”

Light-Mediated 3D Printing of Micro-Pyramid-Decorated Tailorable Wound Dressings with Endogenous Growth Factor Sequestration for Improved Wound Healing

“…A skin substitute that promotes migration of cells involved in skin repair may lead to faster and higher-quality wound closure. Hence, the tip of each pyramid was around 150 μm so that there is some area for the cells to adhere, and the tapered geometry provided a gradient to enhance the cell migration , (Figure S4). To check the stability of the micro-pyramids, the dressings were immersed in PBS for 7 days.…”