scholarly journals A Microfluidic Chip Architecture Enabling a Hypoxic Microenvironment and Nitric Oxide Delivery in Cell Culture

Micromachines ◽  
2020 ◽  
Vol 11 (11) ◽  
pp. 979
Author(s):  
Samineh Barmaki ◽  
Daniela Obermaier ◽  
Esko Kankuri ◽  
Jyrki Vuola ◽  
Sami Franssila ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Cell Culture ◽  
Microfluidic Chip ◽  
Nitric Oxide Donor ◽  
Microfluidic Channels ◽  
Low Oxygen ◽  
Oxygen Scavenger ◽  
Hek 293 ◽  
Human Embryonic Kidney Cells ◽  
Nitric Oxide Delivery

A hypoxic (low oxygen level) microenvironment and nitric oxide paracrine signaling play important roles in the control of both biological and pathological cell responses. In this study, we present a microfluidic chip architecture for nitric oxide delivery under a hypoxic microenvironment in human embryonic kidney cells (HEK-293). The chip utilizes two separate, but interdigitated microfluidic channels. The hypoxic microenvironment was created by sodium sulfite as the oxygen scavenger in one of the channels. The nitric oxide microenvironment was created by sodium nitroprusside as the light-activated nitric oxide donor in the other channel. The solutions are separated from the cell culture by a 30 µm thick gas-permeable, but liquid-impermeable polydimethylsiloxane membrane. We show that the architecture is preliminarily feasible to define the gaseous microenvironment of a cell culture in the 100 µm and 1 mm length scales.

scholarly journals Analysis of the Neuroprotective Effects of Various Nitric Oxide Donor Compounds in Murine Mixed Cortical Cell Culture

2008 ◽  
Vol 72 (5) ◽  
pp. 1843-1852 ◽  
Author(s):  
Aniruddha S. Vidwans ◽  
Sungmee Kim ◽  
Deborah O. Coffin ◽  
David A. Wink ◽  
Sandra J. Hewett
Keyword(s):  
Nitric Oxide ◽  
Cell Culture ◽  
Cortical Cell ◽  
Nitric Oxide Donor ◽  
Neuroprotective Effects

The impact ofN-nitrosomelatonin as nitric oxide donor in cell culture experiments

2008 ◽  
Vol 45 (4) ◽  
pp. 489-496 ◽  
Author(s):  
Utta Berchner-Pfannschmidt ◽  
Suzan Tug ◽  
Buena Trinidad ◽  
Maria Becker ◽  
Felix Oehme ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Cell Culture ◽  
Nitric Oxide Donor ◽  
The Impact

scholarly journals Natriuretic Peptides and Nitric Oxide Stimulate cGMP Synthesis in Different Cellular Compartments

2006 ◽  
Vol 128 (1) ◽  
pp. 3-14 ◽  
Author(s):  
Leslie A. Piggott ◽  
Kathryn A. Hassell ◽  
Zuzana Berkova ◽  
Andrew P. Morris ◽  
Michael Silberbach ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Guanylyl Cyclase ◽  
Cyclic Nucleotide ◽  
Phosphodiesterase Inhibitor ◽  
Soluble Guanylyl Cyclase ◽  
Nitric Oxide Donor ◽  
Dependent Manner ◽  
Channel Activity ◽  
Hek 293 ◽  
Cng Channel

Cyclic nucleotide-gated (CNG) channels are a family of ion channels activated by the binding of cyclic nucleotides. Endogenous channels have been used to measure cyclic nucleotide signals in photoreceptor outer segments and olfactory cilia for decades. Here we have investigated the subcellular localization of cGMP signals by monitoring CNG channel activity in response to agonists that activate either particulate or soluble guanylyl cyclase. CNG channels were heterologously expressed in either human embryonic kidney (HEK)-293 cells that stably overexpress a particulate guanylyl cyclase (HEK-NPRA cells), or cultured vascular smooth muscle cells (VSMCs). Atrial natriuretic peptide (ANP) was used to activate the particulate guanylyl cyclase and the nitric oxide donor S-nitroso-n-acetylpenicillamine (SNAP) was used to activate the soluble guanylyl cyclase. CNG channel activity was monitored by measuring Ca2+ or Mn2+ influx through the channels using the fluorescent dye, fura-2. We found that in HEK-NPRA cells, ANP-induced increases in cGMP levels activated CNG channels in a dose-dependent manner (0.05–10 nM), whereas SNAP (0.01–100 μM) induced increases in cGMP levels triggered little or no activation of CNG channels (P < 0.01). After pretreatment with 100 μM 3-isobutyl-1-methylxanthine (IBMX), a nonspecific phosphodiesterase inhibitor, ANP-induced Mn2+ influx through CNG channels was significantly enhanced, while SNAP-induced Mn2+ influx remained small. In contrast, we found that in the presence of IBMX, both 1 nM ANP and 100 μM SNAP triggered similar increases in total cGMP levels. We next sought to determine if cGMP signals are compartmentalized in VSMCs, which endogenously express particulate and soluble guanylyl cyclase. We found that 10 nM ANP induced activation of CNG channels more readily than 100 μM SNAP; whereas 100 μM SNAP triggered higher levels of total cellular cGMP accumulation. These results suggest that cGMP signals are spatially segregated within cells, and that the functional compartmentalization of cGMP signals may underlie the unique actions of ANP and nitric oxide.

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