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Posts Tagged ‘cellular’

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Poldip2 governs cellular metabolism under hypoxic conditions

Until recently, it was understood that the addition of the prosthetic group lipoate is essential to the activity of critical mitochondrial catabolic enzymes; however, its regulation was unknown. In their paper “Poldip2 is an oxygen-sensitive protein that controls PDH and αKGDH lipoylation and activation to support metabolic adaptation in hypoxia and cancer“, Paredes et al. prove that the Poldip2-controled lipoylation of αKDH and PDH is regulated under hypoxia and in cancer cells, most notably in triple negative breast cancer cells (TNBC).

Using a Don Whitley Scientific  H35 Hypoxystation to establish accurate hypoxic environments, the authors were able to manipulate Poldip2 expression, allowing them to make substantial contributions to the understanding of Poldip2’s essential role in metabolic function during both normoxic and hypoxic conditions. Additionally, they postulate that Poldip2 deficiency may specifically impact the fate and availability of Ac-CoA due to impaired TCA cycle function, potentially providing cancer cells with an advantage for cell proliferation or hypertrophy. Further research will require investigating the interplay between Poldip2 and HIF-1α as well as the regulation of Poldip2 expression at different oxygen concentrations – a task easily achievable thanks to the Hypoxystation’s intuitive interface for effortlessly adjusting the gas composition.

The Whitley  Hypoxystation provides the user with incredible flexibility as it can control oxygen down to 0.1% while providing a temperature and humidity controlled environment with ample working space. The atmosphere is constantly monitored and adjusted by the real-time feedback system to ensure accuracy.

 

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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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