The viability of cells exposed to particle-free air was tested and compared to incubator controls. The ALI cell cultures in the incubator controls were prepared on the same cell culture platform as the ones used in the air exposure; however, they were not removed from the incubator until the toxicity analysis. The particle-free clean air for exposure was generated by filtering laboratory air with a high-efficiency particulate arresting (HEPA) filter. For each exposure, three A549 ALI cell cultures were placed in the exposure dish with 250 μL of media in the basal well. The cell cultures were exposed to clean air regulated by a rotameter attached to a vacuum pump for up to 20 minutes and then returned to the cell incubator for 24 hours before basal media was removed for analysis. Operation flow rates of 4 and 6 LPM were tested in DAVID due to their reliability in deposition in comparison to 8 LPM. The lactate dehydrogenase (LDH) cytotoxicity assay (Thermo Scientific, Waltham, Massachusetts, USA) was used following manufacture protocol to evaluate the cellular effects of the flow rate.
Lactate dehydrogenase ldh cytotoxicity assay
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany
The Lactate dehydrogenase (LDH) cytotoxicity assay is a colorimetric assay that quantifies the amount of LDH released from damaged cells. LDH is a stable cytoplasmic enzyme that is released upon cell lysis or damage. The LDH cytotoxicity assay measures the LDH activity in the culture medium, which is proportional to the number of damaged cells.
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Lab products found in correlation
3 protocols using lactate dehydrogenase ldh cytotoxicity assay
1
Air Exposure Effects on Cellular Viability
2
Evaluating FeOx Nanoparticle Cytotoxicity
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Cells were plated in 96-well plates at 4 × 103 cells per well in serum-containing media and cultured in a 5% CO2 incubator at 37 °C. After 24 h, cells were treated with different concentrations of the FeOxNPs or FeOx-mix (0.02, 0.1, 0.5, 2.5, 10, 50 and 200 μg/mL) at 37 °C for 24 h. CCK-8 cell viability assay was performed according to manufacturer’s protocol (Sigma-Aldrich). Cell viabilities in triplicate wells were measured as the absorbance (450 nm) of reduced WST-8 2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt) using an Infinite 200Pro microplate reader (Tecan Trading AG, Switzerland). Cytotoxicity was also assessed using the Lactate Dehydrogenase (LDH) Cytotoxicity Assay (Thermo Fisher Scientific, Loughborough, UK) according to the manufacturer’s protocol. Cells were prepared and treated with FeOxNPs as described above. FeOxNPs cytotoxic effects in triplicate wells were measured at 492 nm and 690 nm wavelengths, respectively, using a Multiskan Multisoft Primary EIA microplate photometer.
3
Cytotoxicity Evaluation of Hydrogen Peroxide
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The Lactate dehydrogenase (LDH) cytotoxicity assay (Thermo Fisher Scientific, Braunschweig, Germany) was used according to the manufacturer’s protocol in order to analyse cell viability. Fibroblasts were grown under standard conditions in 96 well plates at a density of 15,000 cells per well. Cells were treated with 5 mM hydrogen peroxide (H2O2) for 4 h at 37 °C. Colorimetric measurements were performed on the Microplate Cytation 5 M Cell imaging Multi Mode Reader (Bio-Rad laboratories GmbH, Munich, Germany).
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