Analysis of intercellular communication by flexible hydrodynamic gating on a microfluidic chip
- 592 Downloads
Intercellular Ca2+ waves are propagation of Ca2+ transients among cells that could be initiated by chemical stimulation. Current methods for analyzing intercellular Ca2+ waves are difficult to realize localized chemical stimulations upon the target cell without interfering with adjacent contacting cells. In this paper, a simple and flexible microfluidic method was developed for investigating the intercellular communication of Ca2+ signals. A cross-patterned microfluidic chip was designed and fabricated with polydimethylsiloxane as the structural material. Localized chemical stimulation was achieved by a new strategy based on hydrodynamic gating technique. Clusters of target cells were seeded at the location within 300 μm downstream of the intersection of the cross-shaped microchannel. Confined lateral molecular diffusion largely minimized the interference from diffusion-induced stimulation of adjacent cells. Localized stimulation of the target cell with adenosine 5′-triphosphate successfully induced the propagation of intercellular Ca2+ waves among a population of adjacent contacting cells. Further inhibition studies verified that the propagation of calcium signals among NIH-3 T3 cells was dependent on direct cytosolic transfer via gap junctions. The developed microfluidic method provides a versatile platform for investigating the dynamics of intercellular communications.
KeywordsMicrofluidic chip Hydrodynamic gating Intercellular communication Ca2+ waves
The authors gratefully acknowledge the financial supports from National Basic Research Program of China (2011CB910403) and National Natural Science Foundation of China (30970692 and 21075045).
Numerical simulations of different gating and injection states (MPG 3090 kb)
Visualization study of different gating and injection states (MPG 2756 kb)
- 24.Lee PJ, Hung PJ, Shaw R, Jan L, Lee LP (2005) Appl Phys Lett 86Google Scholar
- 34.Paemeleire K, Martin PE, Coleman SL, Fogarty KE, Carrington WA, Leybaert L, Tuft RA, Evans WH, Sanderson MJ (2000) Mol Biol Cell 11:1815–1827Google Scholar