Primary human AT1 cells were grown in Prigrow III medium (both from Applied Biological Materials, Richmond, BC, Canada) with 10% heat inactivated fetal bovine serum and 1% penicillin-streptomycin in extracellular matrix-coated flasks incubated at 37°C in 95% air-5% CO2 × 12h. About 50,000 cells were seeded in six-well plates and exposed to either air or 95% O2 with 5% CO2 in a sealed humidified chamber (Billups-Rothenberg, Del Mar, CA) at 37°C × 24h.
Sealed humidified chamber
Manufactured by Embrient Inc
The Sealed Humidified Chamber is a laboratory equipment designed to maintain a controlled humidity environment. It features an airtight enclosure with integrated humidification capabilities to regulate the moisture level within the chamber.
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Lab products found in correlation
2 protocols using sealed humidified chamber
1
Primary Human AT1 Cell Hypoxia Exposure
2
Ischemia-Induced Muscle Cell Injury
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The C2C12 cells, maintained in DMEM supplemented with 10% fetal bovine serum (FBS) and 0.1% penicillin/streptomycin, were allowed to differentiate by replacing the FBS with 2% horse serum. Differentiated cells were pretreated with ODC (50–500 μM), carnosine (1 mM) or methylcarcinine (MC, 1 mM) for 8–12 h. To simulate ischemia, differentiated C2C12 cells were subjected to hypoxia for 5, 10, 15, 30 min, and 4 h in Esumi lethal ischemia medium for glucose and nutrient deprivation containing mM: 117 NaCl, 0.9 CaCl2, 12 KCl, 0.49 MgCl2, 4 HEPES, 20 sodium lactate and 5.6 L -glucose; pH 7.4 (Ngoh et al., 2008 (link)). For hypoxia, the C2C12 cells were placed in a sealed humidified chamber (Billups-Rothenberg Inc.) replacing oxygen with 95% N2 and 5% CO2. Cells were harvested in lysis buffer for Western blot and quantitative PCR analysis.
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