In this paper, a simple and versatile method is presented which enables detection of nitric oxide (NO) released from
vascular endothelial cells (ECs) cultured in microfluidic structures. The culturing system and NO measurement
method allow cell shape to be controlled in a non-invasive manner using microfluidic structures while NO release is
monitored for cell shape versus function studies. The culturing system consists of arrays of polydimethylsiloxane
(PDMS) fluidic channels 120 micrometers in depth and ranging from 100 micrometers to 3 mm in width. The
number of channels in each array is varied to yield a constant cell culture surface area (75 mm2) independent of
channel width. The channel surfaces are collagen-coated and ECs are cultured to confluence within the channels. A
cell scraper is then used to scrape extraneous cells cultured between channels, and NO measurements are made 18 to
24 hours later. A chemiluminescence-based sensor system (NOA 280i, Sievers NO Analyzer) is utilized to measure
sample NO. Initial results indicate that NO concentrations can be measured from different microfluidic channel-containing
samples using this method. It is shown that there is no significant difference in NO concentration derived
from channels of different widths even though the degree of cell elongation varies due to physical constraint by
microfluidic channel walls. However, cells treated with TNFα release more NO than untreated cells in fluidic
channels, which is comparable to the function of ECs cultured in conventional culturing systems such as culturing
dishes.
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