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Archive for August, 2016

Jim Uniacke

Physiological Oxygen is Healthier for Cell Cultures

Drs. Timpano and Uniacke, Hypoxystation users at University of Guelph in Ontario, have published a very thorough study examining the molecular basis of cells’ reactions to differing levels of hypoxia. In their paper “Human Cells Cultured Under Physiological Oxygen Utilize Two Cap-binding Proteins to Recruit Distinct mRNAs for Translation” (Journal of Biological Chemistry 291:20; 2016), they examine 2 different translation initiation proteins, eiF4E and eiF4E2, that are activated under either high (>8% O2) or low (<1% O2) oxygen levels, with the aid of mTORC1 or HIF-2α, respectively, and activated simultaneously in an area of low- to mid-level physioxia (1-8% O2). Timpano and Uniacke were able to stably and accurately create low oxygen in their Hypoxystation by Hypoxygen, which provides a closed workstation environment that enables researchers to culture and manipulate cells inside the chamber through gloveless sleeves, eliminating the negative consequences of spikes of higher oxygen and lower temperatures encountered in an incubator as cell cultures are growing. Their research into translational modulation of the proteome using the Hypoxystation gives seminal insights into physioxia as the natural condition for cells, both in vitro and in vivo.

“Culturing cells in ambient air could be far from physiological with resepct to oxygen. Oxygen is a surprisingly neglected factor (in cell culture)” – Dr Timpano and Dr Uniacke, University of Guelph, Ontario, Canada

Through polysome association experiments with cells growing at ambient air versus lower oxygen levels of 1%, 3%, 5%, and 8%, RNA analysis, and m7-GTP cap-binding assays, Timpano and Uniacke were able to demonstrate that the oxygen concentration in the workstation was sufficient to either repress or increase the activity of eiF4E and eiF4E2, reflecting mechanisms that occur during development but also during tumor progression and in ischemic diseases. Cells can reversibly cycle between utilisation of the eiF4E protein, which preferentially binds to the 5′ TOP mRNA’s at >8% O2 and is impaired at hypoxia, and eiF4E2, which is active at <1% O2 and utilizes binding motifs in the 3′ UTR of the mRNA. The eIF4E type of mRNA’s code for housekeeping proteins while the eIF4E2-dependant mRNA’s encode signaling proteins needed to respond to environmental signals, allowing cells to control translation dynamically and giving cancer cells an edge during tumor progression, as hypoxia increases.

Read more

Tell DWS how oxygen makes a difference to your cell culture and win a Don Whitley Scientific power bank to keep your phone charged when you’re on the go! We’ll draw 5 winners from among all entrants (this offer will run for a limited time only).

screengrab

New Video – An Introduction to Don Whitley Scientific

Don Whitley Scientific is delighted to present a new corporate video showing some of the people and processes that have allowed the organisation to remain successful after 40 years in business.

This short video, produced by local company Rejuvenate Productions, gives an insight into how Don Whitley Scientific designs, manufactures and services equipment for laboratories throughout the world. Don Whitley Scientific is very proud of its “one-stop-shop” concept, something that is very much appreciated by customers.

The video starts in the research and development office where members of staff are designing new products and developing ideas for existing products. From there an insight is given into our CNC machine shop where key components are manufactured. The video progresses through the key stages of production, sales, installation and servicing.

We hope we have been able to provide a flavour of how we work with our customers to bring new and innovative solutions to the scientific community.

Whitley H35 Hypoxystation

DWS Goes For Gold at VCCRI Meeting

Don Whitley Scientific Pty will be gold sponsors for this year’s Victor Chang 17th International Symposium at the Garvan Auditorium in Darlinghurst, Sydney. The meeting will incorporate the annual cardiac theme meeting of Stem Cells Australia and the annual meeting of The Australian Network of Cardiac and Vascular Developmental Biologists.

The 2016 event is a series of short lectures on relevant topics by leading international cardiac doctors and researchers. The programme covers a wide range of topics from basic science to clinical medicine. Some of the topics that will be discussed include heart development and congenital heart disease, genetic basis of dilated cardiomyopathy, cardiac muscle function and regeneration, stem cells and new heart failure therapies. For more information visit the VCCRI website.

The team from Don Whitley Scientific will have a trade display and will be able to discuss the wide range of Don Whitley Scientific equipment. Product demonstrations can also be arranged. The VCCRI currently houses two Whitley H35 Hypoxystations which are contributing to fantastic research projects. The Whitley Hypoxystation range offers the ability to work at specific atmospheric conditions, where the environment needs to be physiologically relevant to the levels of oxygen that would be found in the human body. These hypoxic workstations are available in different sizes with a number of options that can be applied to better suit the user’s application, such as front and side loading letterboxes that allow the user to quickly introduce samples into their working area.

The Victor Chang Cardiac Research Institute (VCCRI) was opened in 1994. Victor Chang was a surgeon, researcher and humanitarian who established the Cardiac Transplant Ward and Cardiac Diagnostic Unit at St Vincent’s Hospital in Sydney. Following his untimely death in 1991, the Victor Chang Cardiac Research Institute was established, initially under St Vincents Hospital. It became an independent research facility the following year in 1995. In 1996 it moved to its current home, the Garvan Building, where its premises were opened by the late Diana, Princess of Wales.

 

Scientist Working in Whitley Workstation

Examining the Role of Autophagy in Hypoxic Tumours

Hypoxystation users Tan et al. at the University of Toronto published a paper in June examining the significance of autophagy in cancer development (“Role of Autophagy as a Survival Mechanism for Hypoxic Cells in Tumors“, Neoplasia (2016) 18, 347-355). Autophagy as a means of recycling cell components is induced under stress conditions such as hypoxia, and Tan et al. investigated the correlation of hypoxia and autophagy in solid tumours in the context of resistance to cancer therapeutics.

Cells were cultured in the H35 Hypoxystation for up to 48 hours at hypoxia (0.2 %) and compared to cells grown at ambient oxygen level. Gene silencing of autophagy proteins ATG7 and BECLIN1 with shRNA resulted in decreased cell survival under hypoxia, and inhibition of autophagy with pantoprazole exacerbated the loss of viability in the knock-down cells under hypoxia, demonstrating the cyto-protective effects of these autophagy proteins. Using the Seahorse XFe Analyzer to assess oxygen consumption in wild-type and silenced cells, Dr. Tan’s lab found reduced respiration when autophagy is disrupted, possibly due to accumulation of dysfunctional mitochondria in these mutant cells. The H35 Hypoxystation  Dr. Tan’s lab used for these studies creates a closed environment with controlled temperature, humidity, CO2 and oxygen, in which cells are cultured and manipulated over the course of days and weeks without the need to transfer into ambient conditions. The combination of an Hypoxystation and an i2 Instrument Workstation is designed to accommodate the specific requirements of the Seahorse XFe Analyzer for the duration of the metabolism assays investigating oxygen consumption and extracellular acidification.

Please visit the Don Whitley Scientific website for more published papers featuring the Whitley Workstation range or contact us to discuss your hypoxia needs.

Thuy Do pressing touchscreen

New Option – Catalyst Monitoring System

This new facility is a very efficient and effective way of alerting users to both the condition of the catalyst and the anaerobic atmosphere in a Whitley Workstation. This patented application is an ingenious software based system, which requires only a minimal amount of unobtrusive hardware.

The Catalyst Monitoring System is a factory-fitted option for Whitley Anaerobic Workstations. It is set to conduct the test overnight so that when you arrive at the lab in the morning, a pop-up box on the touchscreen informs you of the status of the catalyst. ‘Green’ means all is fine and there is plenty of life left in the catalyst; ‘amber’ means the catalyst performance is reduced; and ‘red’ means that the catalyst should be changed to maintain good anaerobic conditions.

The system also allows for the test to be run manually if required. This test is very valuable because it confirms that the catalyst is working and that hydrogen is present in the atmosphere, both of which are essential to maintain strict anaerobic conditions.

This option can be used either on its own or in conjunction with the Whitley Anaerobic Conditions Monitor, which provides continuous monitoring of the level of oxygen present in your workstation.

 

A07227 - Catalyst monitoring system