Following our recent example (30 (link)), we screened the activity of our PS family in hypoxia (1% O2) to evaluate oxygen independence relative to a compound known to be highly oxygen-dependent for its activity [Ru(bpy)2(dppn)]Cl2 (60 (link), 61 (link)). This was performed 4 days after the normoxic screen. Hypoxic cell culture was conducted inside a Biospherix Xvivo X3 at 1% O2 (5% CO2, 37 °C). Following cell adhesion post-seeding (2–3 h) in normoxia, plates were incubated at 1% O2 for 2–3 h before dispensing compound dilutions. After an overnight incubation in hypoxia, plates to be irradiated were covered with film (VWR, 89134–428) to maintain the low oxygen environment during light treatment. Following light treatment, films were removed and all low oxygen plates were replaced to the normoxic incubator and incubated overnight before final viability measurements.
Xvivo x3
Manufactured by Biospherix
Sourced in United States
The Xvivo X3 is a laboratory equipment designed for maintaining and monitoring cell cultures in a controlled environment. It provides precise temperature, humidity, and gas regulation to support the growth and preservation of cells. The core function of the Xvivo X3 is to create a stable, physiologically relevant in vitro setting for cell-based research and applications.
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Lab products found in correlation
2 protocols using xvivo x3
1
Evaluating Hypoxic Activity of PS Compounds
2
Cisplatin Sensitivity in Hypoxic Pten Cells
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To assess Pten/WT and Pten/KO cells’ susceptibility to cisplatin treatment in hypoxia (1% pO2), cells were seeded in 96-well plates under standard oxygen conditions and allowed to adhere to the culture surface. After 24 h, the medium was changed to a pre-equilibrated hypoxic medium, and the cells were cultured in an XVivo X3 workstation (Biospherix, USA) in 1% pO2 and 5% CO2 at 37 °C for a further 24 h. Next, cisplatin was added at the final testing concentrations, as previously used in normoxic conditions, and Alamar Blue was used, as described above. The conditioned media were collected from cells cultured in hypoxia for 72 h without drugs, and PAI-1 secretion was detected using a Mouse PAI-1 DuoSet ELISA (R&D Systems, USA), as described above.
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