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Posts Tagged ‘HIFs’

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Culturing Cells in Ambient Air is Far From Physiological

Based on the premise that the physiological range of oxygen in tissues is between 1- 8%, and pathologies from cancer to diabetes are characterised by much lower oxygen levels, researchers worldwide are cultivating their cell cultures in the Hypoxystation by Don Whitley Scientific. The Hypoxystation provides physiologically relevant conditions for cell culture and manipulation to ensure authentic behaviour of cells. User-defined parameters for temperature, CO2, O2 and humidity, plus the workstation format, where cells reside throughout the entire duration of the assays, minimise the extra-physiologic oxygen shock that is known to negatively impact cell metabolism and growth.

Numerous recent publications by our Hypoxystation users demonstrate that cell culture conditions which mimic physioxia, are essential in avoiding the significantly impaired growth rates, reduced lifespan, and altered molecular behaviour encountered in cells cultivated at ambient conditions. Oxygen levels in tissues are in constant flux; they change in response to functional status and blood delivery in the organs, and this too can be re-created in the Hypoxystation through programmed oxygen profiling.

Recent research using the Hypoxystation to investigate hypoxia inducible factors and the array of signalling pathways that regulate angiogenesis, metabolism, redox homeostasis, inflammation, and cell death, and the many other processes which enable the cellular and organismal response to hypoxia, “highlights the importance of oxygen as a cell culture parameter when making physiological inferences” (Timpano and Uniacke, 2016).

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From: Burr et al. (2016) “Mitochondrial Protein Lipoylation and the 2-Oxoglutarate Dehydrogenase Complex Controls HIF1a Stability in Aerobic Conditions” Cell Metabolism 25, 740–752


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Hypoxystation In The Lab – Sequestration In Speckles

The HIF (hypoxia-inducible factor) family of oxygen-sensing proteins are a crucial element of cells’ responses to alterations in their immediate environment, kicking off a signaling cascade involving more than 1000 genes. Hypoxystation users Taylor and See at the University of Liverpool describe novel insights into the subcellular localization of some of the HIF proteins and why the “where” determines the “how”.

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HIF-2α and HIF-1α both form heterodimers with HIF-1β, and while similarities abound between the isoforms, the two subunits are differentially expressed and regulated and have distinctly separate target genes. Taylor and See triggered HIF activation using microscope stage incubators and the Hypoxystation by Don Whitley Scientific to incubate HeLa cells in hypoxia (1%). They found that while HIF-1α distributes homogenously in the nucleus, HIF-2α diffuses freely through the nucleus but is concentrated in speckles that are tethered to nuclear structures close to active RNA polymerases. This distribution is not significantly altered by low oxygen levels.

The Hypoxystation provides physiologically relevant, in vivo conditions for cell culture and manipulation to ensure authentic behavior of cells. User-defined parameters for temperature, CO2, O2 and humidity, plus the workstation format, where cells reside throughout the entire duration of the assays, minimize the extra-physiologic shock that is known to negatively impact metabolism and growth. As the degree and duration of hypoxia are among the factors controlling HIF activity, the customizable oxygen atmosphere inside the Hypoxystation can contribute to deciphering the functionality of oxygen-sensitive signaling pathways.