Cells were exposed to hypoxia by incubating at 37 °C in 1% O2/5% CO2/94% N2 anaerobic condition in an air-tight modular incubator chamber (Billups-Rothenberg Inc., San Diego, CA, USA).46 (link)
Airtight modular incubator chamber
Manufactured by Embrient Inc
Sourced in United States
The Airtight modular incubator chamber is a laboratory equipment designed to provide a controlled environment for various applications. It features an airtight construction and modular design, allowing for customization and adaptability to specific experimental needs.
Automatically generated - may contain errors
Lab products found in correlation
Gas analyzer, by GE Healthcare (2 mentions)
Aestiva 5, by GE Healthcare (1 mentions)
Desflurane, by Baxter (1 mentions)
Dynabeads human t activator cd3 cd28, by Thermo Fisher Scientific (1 mentions)
Hepes buffer solution, by Thermo Fisher Scientific (1 mentions)
Cell culture insert, by Greiner (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Cellstar, by Greiner (1 mentions)
7 protocols using airtight modular incubator chamber
1
Hypoxia Exposure Protocol
2
Desflurane Exposure on Cultured Neurons
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After 1-h incubation at 37 °C and 5% CO2, the cells were exposed to 0.5 minimum alveolar concentration (MAC) equivalent of desflurane (4.3 Vol%; Baxter Corporation, Mississauga, ON, Canada) in an airtight modular incubator chamber (Billups-Rothenberg) for one hour to mimic the inhalation. desflurane-medical air gas mixtures were vaporized using a Datex-Ohmeda Aestiva/5 vaporizer and concentrations were monitored with a GE Healthcare Gas Analyzer. Controls were exposed to medical air only (79% Nitrogen, and 21% Oxygen; Air Liquide). After 1 h of desflurane-medical air mixture or just medical air exposure, the neurons were placed back and maintained in an incubator (37 °C, 5% CO2) until use. This exposure time was consistent with previous studies, aimed at optimal gas exchange with minimum out of incubator time for culture neurons.
3
Sevoflurane Exposure on Neurons
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After 1 h incubation at 37 °C and 5% CO2, the cells were exposed to approximately 0.5 minimum alveolar concentration (MAC) equivalent (for neonates) of sevoflurane (1.6%) in an airtight modular incubator chamber (Billups-Rothenberg, San Diego, CA, USA) for one hour. Sevoflurane-medical air–oxygen gas mixtures (21% O2) were then vaporized using a Datex-Ohmeda Aestiva/5 vaporizer and concentrations were monitored with a GE Healthcare Gas Analyzer. Controls were exposed to medical air only. After 1 h sevoflurane-medical air–oxygen mixture or just medical air exposure, the neurons were placed back and maintained in an incubator (37 °C, 5% CO2) until use.
4
Anakinra Modulates GBM-PBMC Crosstalk
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GBM cells were seeded in six well plates (Cellstar®, Greiner Bio-one, Austria) containing 2 mL DMEM. After adherence, medium was changed to 3 ml RPMI 1640 medium (Gibco), supplemented with 10% FCS, 1% L-Glutamine, 1% penicillin/streptomycin, and 1% HEPES buffer solution (Gibco), and PBMCs, stimulated with Human T-Activator CD3/CD28 Dynabeads (Thermo Fisher, Waltham, MA, USA) were added reaching a final GBM cell–PBMC ratio of 1:10. Cells were incubated in the present or absence of anakinra (1 µg/mL) under moderate hypoxic conditions in an airtight modular incubator chamber (Billups-Rothenberg, San Diego, USA) at 5% O2, 40 mmHg CO2 and 37 °C. For indirect co-culture experiments, GBM cells were separated from PBMCs by cell culture inserts (pore diameter: 0.4 μm) (Greiner Bio-one, Austria). After 24 h or 48 h cells, were harvested for further analyses. Cell culture supernatant was aliquoted and stored at −80 °C. Supernatant from direct co-cultures were subsequently subjected to native GBM cells, seeded the previous day. After addition of anakinra, cells were also incubated in moderate hypoxia for 48 h.
5
Hypoxia Induction for Cell Assays
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Control cells were incubated for 6 or 24 h under normoxic conditions (21% O2, 5% CO2 and 74% N2 at 37°C) in a humidified incubator. According to the manufacturer’s protocols, hypoxic conditions (termed hypoxia) were induced using an airtight modular incubator chamber (Billups-Rothenberg, Inc., San Diego, CA, USA). Briefly, the cells [1×104 cells/well in cell viability assay; 2×104 cells/well in tube formation assay; and 1×105 cells/well in western blotting, ELISA and flow cytometry (FCM) assays] were sealed in the modular incubator chamber with a sterile 1X PBS reserve to maintain humidity, and then purged with a reduced O2 gas mixture (1% O2, 5% CO2 and 94% N2) at 37°C for 6 h or 24 h.
6
Exploring Preterm and Term WJ-MSCs Under Hyperoxia
7
Hypoxia Exposure Viability Assay
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All hypoxic experiments were performed in an airtight modular incubator chamber (Billups-Rothenberg), which was deoxygenated by positive infusion of 2% O 2 in a CO 2 -nitrogen gas mixture for 5 minutes. Cultures were then placed in a standard humidified tissue incubator. There were no statistically significant differences in viability by crystal violet or trypan blue exclusion (data not shown). Parallel cultures were placed in normal conditions (20% O 2 ) for all time points. Cells were harvested after 24 hours. All experiments were performed in triplicate.
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