Weanling C57BL/6 male mice (n = 4–5/group) were given free access to Harlan AIN-93G diets containing iron at 4 mg/kg (TD.10210) for an FeD diet or 35 mg/kg (TD.10211) for an FeA diet for 14 weeks (by weight 17.7% protein, 60.1% carbohydrate, and 7.2% fat; Harlan Teklad, Madison, WI). Weanling mice were fed the same diets for 10 weeks and then transferred into a hypoxia chamber and exposed to 8% O2 for 4 weeks with continuation of the same diets. The experiment design is summarized in Fig. 1 . Oxygen levels were regulated by a ProOx 110 oxygen controller (BioSpherix). The chamber was opened for ∼10 min every 3 days for maintenance and refeeding. At the end of the studies, animals were anesthetized with isoflurane, and blood collection was performed by cardiac puncture. Animal protocols were approved by the University of Utah and Wake Forest University Institutional Animal Care and Use Committees.
Proox 110 oxygen controller
Manufactured by Biospherix
Sourced in United States
The ProOx 110 is an oxygen controller designed to precisely regulate oxygen levels in a variety of laboratory applications. It features digital displays for monitoring and adjusting oxygen concentrations. The device is capable of delivering a controlled oxygen environment, but its specific intended uses are not provided.
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Lab products found in correlation
6 protocols using proox 110 oxygen controller
1
Dietary Iron Modulation in Hypoxic Mice
2
Baicalin Treatement in Oxygen-Induced Retinopathy
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ICR pups were randomly divided into 4 groups: a normoxia group, an oxygen-exposed group (OIR group), and 2 groups that received different doses of baicalin. Each group had 1 nursing mother and 5-7 pups, and all experiments were repeated 3 times. At least 15 pups were included in each group. Oxygen-induced retinopathy was induced in ICR pups using previously published methods (14) (link). For the OIR model, the newborn pups at post-natal day 7 (P7), along with their mothers, were transferred to a chamber supplied with 75 ± 2% oxygen, which was under continual monitoring with a ProOx 110 oxygen controller (Biospherix, Ltd., Lacona, NY, USA) for 120 h. On P12, the mice were returned to an atmosphere of normal room air and were given daily intraperitoneal (IP) injections of 10% DMSO in water (vehicle) or baicalin dissolved in vehicle at 1 mg/kg (B1, low-dose) or 10 mg/kg (B10, highdose). The mice in the normoxia group were maintained in normal room air from birth until P17.
3
Hypoxia-Induced Cardiomyocyte Proliferation
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After 24 hours of normal culture, the medium was replaced with F10 medium containing 0.1% FCS, and 0.1 mmol/L BrdU (bromodeoxyuridine; to inhibit the proliferation of fibroblasts) to induce hypoxia. The cultured cardiomyocytes were exposed to hypoxia stimulation in a BioSpherix C‐Chamber (model C‐274; BioSpherix) with a standard cell culture chamber. A concentration of 5% O2 and 5% CO2 was maintained inside the C‐Chamber by injecting N2 and CO2 using a ProOx 110 oxygen controller and a ProCO2 CO2 controller (BioSpherix). For the control group, the C‐Chamber was maintained at 37°C and filled with 5% CO2 and 95% air. After 60 minutes of hypoxia, the cells were then incubated under normal conditions for 3 hours.
4
Neonatal Oxygen Exposure in Mice
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Nursing dams with P7 pups were housed in a Perspex chamber (BioSpherix) and exposed continuously to 74% ± 1% oxygen in air maintained by a ProOx110 oxygen controller (BioSpherix). Duration of oxygen exposure and subsequent recovery time in room air is indicated in each figure. Pups were fostered to BALB/c females following exposure to 3 days of high oxygen to prevent oxygen toxicity in dams.
5
Oxygen-Induced Retinopathy and Sulodexide
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ICR pups were randomly divided into three groups: a normoxia group (control group), an oxygen-exposed group (OIR group), and a sulodexide group; each group had one nursing mother and 5-7 pups. Oxygen-induced retinopathy was induced in ICR pups, as described previously (23) (link). For the OIR model, the newborn pups were transferred at post-natal day (P) 7 along with their mother to a chamber supplied with 75 ± 2% oxygen, under continual monitoring with a ProOx 110 oxygen controller (Biospherix, Ltd., Lacona, NY, USA) for 120 h. On P12, the mice were returned to the room air and given daily intraperitoneal (IP) injections of vehicle (saline) or 5-15 mg/kg of sulodexide dissolved in the vehicle. The mice in the normoxia group were maintained in room air from birth until P17.
6
Tracking RBC Turnover in Hyperoxia
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A hyperbaric chamber (ProOx 110 oxygen controller, BioSpherix, USA) was kindly provided by the laboratory of Dr. Claudio Franco at Instituto de Medicina Molecular (Lisboa). Mice were kept there for 5 days in a hyperoxic atmosphere (75% O2). After that period, mice returned to normoxia conditions. At this moment, two initial biotin injections (1mg/200 μl) stained all the RBCs that were already in circulation before the hyperoxia condition (termed “biotin high RBCs” in the text). Seven days later, a third biotin injection (0.6 mg/200 μl) labeled all the newborn RBCs formed during the hyperoxia treatment (referred to as “biotin intermediate RBCs” in the text). Both populations were monitored once a week for 31 days by flow cytometer (BD LSRFortessa). To that end, 1 μl of blood was collected from the tail of each mouse by poking with a 25g needle, and stained with allophycocyanin (APC) conjugated streptavidin (Biolegend), which binds the biotin.
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