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Article

An Inexpensive Incubator for Mammalian Cell Culture Capable of Regulating O2, CO2, and Temperature

Department of Biological Sciences, Brock University, St. Catharines, ON L2S 3A1, Canada
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Author to whom correspondence should be addressed.
Academic Editor: John T. Hancock
Received: 15 February 2022 / Revised: 1 March 2022 / Accepted: 9 March 2022 / Published: 14 March 2022
(This article belongs to the Special Issue Feature Papers in Oxygen)
Mammalian cell culture is widely used for discovery and development. Recently, increasing attention has been paid to the importance of maintaining physiologically-relevant conditions in cell culture. Although oxygen level is a particularly important consideration, it is rarely regulated by experimentalists. The atmospheric O2 levels commonly used in cell culture are significantly higher than those experienced by most mammalian cells in vivo, leaving cells susceptible to oxidative damage, senescence, transformation, and otherwise aberrant physiology. A barrier to incorporating O2 regulation into most cell culture workflows has been the expense of investing in new equipment, as the vast majority of laboratory CO2 incubators do not regulate O2. Here, we describe an inexpensive (<CAD 1000), portable and user-friendly O2/CO2 incubator that can establish and maintain physiological O2, CO2, and temperature values within their physiological ranges. We used an Arduino-based approach to add O2 and CO2 control to a temperature-regulating egg incubator. Our incubator was tested against a commercial laboratory O2/CO2 incubator. Using Presens OxoDish technology, we demonstrate that at a setpoint value of 5% gas-phase incubator O2, media O2 averaged 5.03 (SD = 0.03) with a range of 4.98–5.09%. MCF7, LNCaP and C2C12 cell lines cultured in the incubator displayed normal morphology, proliferation, and viability. Culture for up to one week produced no contamination. Thus, our incubator provides an inexpensive means of maintaining physioxia in routine mammalian cell culture. View Full-Text
Keywords: oxygen; physioxia; hypoxia; cell culture; incubator oxygen; physioxia; hypoxia; cell culture; incubator
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MDPI and ACS Style

Samokhin, P.; Gardner, G.L.; Moffatt, C.; Stuart, J.A. An Inexpensive Incubator for Mammalian Cell Culture Capable of Regulating O2, CO2, and Temperature. Oxygen 2022, 2, 22-30. https://0-doi-org.brum.beds.ac.uk/10.3390/oxygen2010003

AMA Style

Samokhin P, Gardner GL, Moffatt C, Stuart JA. An Inexpensive Incubator for Mammalian Cell Culture Capable of Regulating O2, CO2, and Temperature. Oxygen. 2022; 2(1):22-30. https://0-doi-org.brum.beds.ac.uk/10.3390/oxygen2010003

Chicago/Turabian Style

Samokhin, Philip, Georgina L. Gardner, Chris Moffatt, and Jeffrey A. Stuart. 2022. "An Inexpensive Incubator for Mammalian Cell Culture Capable of Regulating O2, CO2, and Temperature" Oxygen 2, no. 1: 22-30. https://0-doi-org.brum.beds.ac.uk/10.3390/oxygen2010003

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