Microfluidic platform for the study of intercellular communication via
soluble factor-cell and cell-cell paracrine signaling |
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| Authors: | Matthew B Byrne Lisa Trump Amit V Desai Lawrence B Schook H Rex Gaskins Paul J A Kenis |
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| Institution: | 1.Department of Chemical & Biomolecular Engineering, University of Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA;2.Institute for Genomic Biology, University of
Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA;3.Department of Animal Sciences, University of
Illinois at Urbana-Champaign, Urbana, Illinois 61801, USA |
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| Abstract: | Diffusion of
autocrine and paracrine signaling molecules allows cells to
communicate in the absence of physical contact. This chemical-based, long-range
communication serves crucial roles in tissue function, activation of the immune system,
and other physiological functions. Despite its importance, few in vitro
methods to study cell-cell signaling through paracrine factors are available today. Here,
we report the design and validation of a microfluidic platform that enables (i) soluble molecule-cell and/or
(ii) cell-cell paracrine signaling. In the microfluidic platform, multiple cell
populations can be introduced into parallel channels. The channels are separated by arrays
of posts allowing diffusion of paracrine molecules between cell
populations. A computational analysis was performed to aid design of the microfluidic platform.
Specifically, it revealed that channel spacing affects both spatial and temporal
distribution of signaling molecules, while the initial concentration of the signaling
molecule mainly affects the concentration of the signaling molecules excreted by the
cells. To validate the microfluidic platform, a model system composed of the
signaling molecule lipopolysaccharide, mouse macrophages, and engineered human embryonic
kidney
cells was introduced into the platform. Upon diffusion from the first
channel to the second channel, lipopolysaccharide activates the macrophages which begin to
produce TNF-α. The TNF-α diffuses from the second channel to the third channel to
stimulate the kidney
cells, which express green fluorescent protein (GFP) in response. By
increasing the initial lipopolysaccharide concentration an increase in fluorescent
response was recorded, demonstrating the ability to quantify intercellular communication
between 3D cellular constructs using the microfluidic platform reported
here. Overall, these studies provide a detailed analysis on how concentration of the
initial signaling molecules, spatiotemporal dynamics, and inter-channel spacing affect
intercellular
communication. |
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