Standing surface acoustic wave based cell coculture

Abstract

Precise reconstruction of heterotypic cell-cell interactions in vitro requires the coculture of different cell types in a highly controlled manner. In this article, we report a standing surface acoustic wave (SSAW)-based cell coculture platform. In our approach, different types of cells are patterned sequentially in the SSAW field to form an organized cell coculture. To validate our platform, we demonstrate a coculture of epithelial cancer cells and endothelial cells. Real-time monitoring of cell migration dynamics reveals increased cancer cell mobility when cancer cells are cocultured with endothelial cells. Our SSAW-based cell coculture platform has the advantages of contactless cell manipulation, high biocompatibility, high controllability, simplicity, and minimal interference of the cellular microenvironment. The SSAW technique demonstrated here can be a valuable analytical tool for various biological studies involving heterotypic cell-cell interactions.

Figure 1

(a) Working mechanism of our SSAW-based cell coculture platform. (b) An optical image of our SSAW-based cell coculture device.

Figure 2

Schematics of the SSAW-based cell coculture technique.

Figure 3

(a–e) Micrographs showing culture of patterned HeLa cells in our SSAW-based microfluidic device for up to 24 h. (f) Live/dead staining results indicate that most of the HeLa cells remain viable at the end of 24 h culture (green, live cells; red, dead cells).

Figure 4

Coculture with SSAW-based sequential cell patterning. (a) Mechanism of patterning two types of cells in different positions with the phase-shift approach. (b) Green fluorescent image showing first-seeded HeLa cells. (c) Red fluorescent image showing second-seeded HeLa cells. (d) Merged image showing green and red HeLa cells grown in alternate lines.

Figure 5

Fluorescent images of (a and b) HeLa cell monoculture at 2 and 24 h and (c and d) HeLa and HMVEC-d coculture at 2 and 24 h in our SSAW device.

Figure 6

Quantitative analysis of HeLa cell movement in monoculture and coculture. (a and b) Three typical movement trajectories (in blue, green, and red lines) for HeLa cells in on-chip (a) monoculture and (b) coculture. (c) Average movement path lengths of the tracked HeLa cells plotted against culture time for the four groups. (d and e) Comparison of (d) average movement path length and (e) average distance from origin for the tracked HeLa cells at 12 h among four different groups. The error bars represent the standard deviation (n = 30 for each group; ns, P > 0.05; *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001).