Analysis of intercellular communication by flexible hydrodynamic gating on a microfluidic chip.
Intercellular Ca(2+) waves are propagation of Ca(2+) transients among cells that could be initiated by chemical stimulation. Current methods for analyzing intercellular Ca(2+) 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 Ca(2+) 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 Ca(2+) 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.
Britton Chance Center for Biomedical Photonics at Wuhan National Laboratory for Optoelectronics-Hubei Bioinformatics, Systems Biology Theme, Department of Biomedical Engineering, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, 430074, China.
Peng
P
Chen
Pu
P
Chen
Xiaojun
X
Feng
Wei
W
Du
Bi-Feng
BF
Liu
D000255:Adenosine Triphosphate / Q000737:chemistry; D000818:Animals; D002021:Buffers; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D020013:Calcium Signaling; D002450:Cell Communication; D003198:Computer Simulation; D003600:Cytosol / Q000378:metabolism; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004867:Equipment Design; D017629:Gap Junctions / Q000378:metabolism; D057446:Hydrodynamics; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D044085:Microfluidics; D008956:Models, Chemical; D008962:Models, Theoretical; D041681:NIH 3T3 Cells; D055095:Optics and Photonics; D015398:Signal Transduction; D013314:Stress, Mechanical; D013997:Time Factors
D002021:Buffers; D004129:Dimethylpolysiloxanes; C013830:baysilon; D000255:Adenosine Triphosphate; D002118:Calcium