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Posts Tagged ‘HEPA Filtration’

Whitley Hypoxystation

The Leukemic Stem Cell Niche: Adaptation to “Hypoxia” Versus Oncogene Addiction

Hematopoietic stem cells (HSC) are responsible for constantly maintaining and replenishing the supply of new blood and immune cells. They give rise to both lymphoid and myeloid progenitor cells, which then proceed to differentiate down their respective paths to form various specialized cells such as erythrocytes, macrophages, B and T cells, to name a few. Within the body, HSCs are found to reside in extremely low oxygen environments called stem cell niches (SCN). Like all other regulated cell cycles, HSCs can lead to cancers such as leukemia and lymphoma if cell division becomes uncontrolled.

MEL (a), Kasumi-1 (b), or NB4 (c) cells were incubated in atmosphere at 0.1% O2 and lysed at the indicated times, and total cell lysates were subjected to immunoblotting with the indicated antibodies. GAPDH, H4, or ARD1 were detected to verify loading equalization. Migration of molecular weight markers is indicated on the left (kDa). For each cell population, one out of three independent experiments with similar outcome is shown.

Figure 2: Suppression of oncogenic proteins driving non-CML blood neoplasias in the course of cell “adaptation to hypoxia.” MEL (a), Kasumi-1 (b), or NB4 (c) cells were incubated in atmosphere at 0.1% O2 and lysed at the indicated times, and total cell lysates were subjected to immunoblotting with the indicated antibodies. GAPDH, H4, or ARD1 were detected to verify loading equalization. Migration of molecular weight markers is indicated on the left (kDa). For each cell population, one out of three independent experiments with similar outcome is shown.

In this paper, Cheloni et al. primarily focused on chronic myeloid leukemia (CML).

The authors hypothesized that suppression of the BCR-Abl oncogene is likely a key positive regulator of LSC survival within “hypoxic” SCNs. To analyse the various mechanisms and responses that CML cells demonstrate within SCNs, the authors studied correlations between varied oxygen and glucose concentrations with the amount of BCR-Abl produced. All tests were performed using two human CML cell lines, K562 and KCL22. Testing conditions were precisely controlled to mimic the SCN environment as close as possible. Using a Hypoxystation supplied by Don Whitely Scientific, a water-saturated atmosphere comprising of 0.1% O2, 94.9% N2 and 5% CO2 was generated and maintained.

The research conducted and documented by Cheloni et al. has provided great insight into several key regulatory mechanisms associated with leukemic stem cells as well as an explanation for their notorious reputation for having high relapse rates. Additionally, they established that the triggering of oncogene suppression associated with CML is due to severe energy restriction rather than simply the “adaptation to hypoxia.”

The Don Whitley Scientific Hypoxystation provides the user incredible flexibility as it can control oxygen down to 0.1% while providing a temperature and humidity controlled environment with ample working space. The extreme precision provided by the Hypoxystation was critical to the collection of accurate and reliable data when recreating the demanding environment that comprises SCNs. The atmosphere is constantly monitored and adjusted by the real-time feedback system to ensure accuracy and ISO class 3 clean room HEPA filtration is also available for long term cell culture applications.

Choose your atmosphere with the Hypoxystation hypoxia chamber. Accurately control O2, CO2, Temperature and Humidity.   

Hypoxystation is the only hypoxic chamber purpose built for physiological cell culture research. Specifically designed to create normoxic, hypoxic and anoxic conditions within a controlled and sustained workstation environment, this hypoxic incubator is ideal for research requiring the ability to accurately control O2, CO2, temperature and humidity. The Whitley Internal HEPA Filtration System provides a particle-free internal environment that exceeds ISO 14644 class 3 clean atmosphere.  With such accurate control and the ability to manipulate cells in situ without altering the incubation environment, research into cell biology can be performed over a comprehensive range of oxygen tensions with precision. Don Whitley Scientific offer the following range of Hypoxystations:

                                                                                   

Hypoxystation H35

Hypoxystation H45

Hypoxystation H85

Hypoxystation H135

i2 Instrument Workstation

Application possibilities for Hypoxystation are endless; it is being used for research into tumour microenvironment, hypoxia pathways and HIF signalling, in vitro modelling of in situ environments, cancer cell metastasis, angiogenesis, and many other fields where cells may benefit from a more physiological atmosphere.

 

hepa filter

HEPA Filtration from Don Whitley Scientific

For over forty years, Don Whitley Scientific have utilised experience and expertise to ensure products make working methods more straight forward for our laboratory customers. With more and more importance placed on clean air, HEPA filtration can provide an extra measure to ensure research carried out within Whitley Workstations is done so in an environment that achieves particulate concentrations exceeding the levels detailed in ISO 14644-1 Part 1, Table 1, (Class 3) Classification of Air Cleanliness.

 

Tests have been carried out by Don Whitley Scientific that show just how much of a difference a HEPA filtration system makes to the atmosphere of a Whitley Workstation. Don Whitley Scientific have been manufacturing products for over 40 years and the introduction of HEPA filtration was a logical step to make sure our customers have an option that falls in line with the latest laboratory trends and requirements.

The benefits of the Whitley HEPA filtration system are:

  • The Whitley system utilises an internal HEPA filter. The atmosphere is cleaned quickly and continuously because the entire internal atmospheric volume of every Whitley Workstation passes through the filter hundreds of times an hour.
  • The atmosphere passing through the filter creates a laminar flow-like effect covering around 94% of the width of the chamber.
  • The location of the Whitley HEPA filter inside the workstation ensures it cannot get wet. Therefore it cannot become saturated with moisture and rendered ineffective.

Don Whitley Scientific has worked with one of the world’s leading filter manufacturers to develop a filter specifically tailored to suit the requirements of anaerobic workstations. Adding HEPA filter technology to the workstation means creating an environment where microbiology samples and cellular cultures can grow free from the risk of contamination.

HEPA filtration was introduced as an option after decades of experience in microbiology. We are confident that our thorough approach to product development and product testing has resulted in a HEPA filtration system superior to any other systems fitted to positive pressure modified atmosphere workstations. The video below explains Whitley HEPA filtration in further detail.

 

 


Explore the Whitley HEPA Workstation range


 

A135 HEPA_No sleeves CROPPED

The Whitley A135 HEPA Anaerobic Workstation

Don Whitley Scientific is proud to present the new Whitley A135 HEPA Anaerobic Workstation: the tallest, deepest, widest anaerobic workstation in our extensive range. It is specifically designed to maintain strict anaerobic conditions within a controlled environment.
 
The Whitley A135 HEPA has an enormous capacity of almost 600 litres, perfect for working and incubation. It will accommodate a variety of items of equipment such as microscopes, plate readers and colony pickers. (Be sure to discuss your requirements with us at the earliest opportunity).

With a volume twice that of a Whitley A35 Anaerobic Workstation, the A135 also comes fitted with a large removable front – ideal for the transfer of equipment in and out of the workstation. Other great DWS features include:

•           Anaerobic conditions monitoring system
•           Internal HEPA filtration system with enhanced biological containment
•           Rapid 12 litre airlock for sample transfer
•           7” full colour touch screen, Ethernet-enabled for remote access.

135 features

Internal dimensions (w x d x h): 1100mm x 750mm x 710mm
External dimensions (w x d x h): 1452mm x 1056mm x 993mm

Without compromising quality or precision, this innovative product allows you to take your research to new heights whilst maintaining confidence in your results.

 

For more information – email us at sales@dwscientific.co.uk or call us on +44 (0) 1274 595728

dws-stand

Don Whitley Scientific Exhibit at Key Cardiovascular Meeting

Don Whitley Scientific recently attended the BSCR (British Society of Cardiovascular Research) Annual Meeting, held this year at The University Of Leeds. The focus of the meeting was on “Cardiovascular Disease and Diabetes” and the event featured fantastic talks and poster presentations from renowned figures in Cardiovascular research.

The Don Whitley Scientific stand featured a H45 Hypoxystation. The H45 is a large, three port cell culture workstation, ideal for replicating physiologically relevant atmospheres through the control of oxygen, temperature and humidity. Working in a Hypoxystation means cell cultures and other samples are within a stable atmosphere that matches the specific microenvironments that exist inside the human body, meaning research results are dependable and accurate.

 

At The University of Liverpool, Dr Michael Cross is conducting cardiovascular research using a Whitley H35 HEPA Hypoxystation as he looks at Low Oxygen Drug Toxicity Testing in Cardiac Spheroids

The Whitley Hypoxystation range can be viewed here

i2 and H35 HEPA at ICR London

Whitley i2 Instrument Workstation used in deciphering the metabolic properties of Breast Cancer

The Institute of Cancer Research, London, is Europe’s largest cancer centre and one of the world’s most influential research institutes, with an outstanding record of achievement dating back more than 100 years.

Staff at the ICR with their new Whitley Workstations

ICR staff with their i2 and H35 HEPA Workstations

Cancer cells display unique alterations in their signalling and metabolic circuitry in order to fuel their growth and proliferation and/or adapt to conditions of oxygen and nutrient deprivation. The Signalling and Cancer Metabolism Team, led by Dr George Poulogiannis uses high-throughput technologies including mass spectrometry-based metabolomics, and utilizes the Seahorse XF Analyzer to measure the basal oxygen consumption and glycolysis rates in order to reveal the metabolic dependencies of breast cancer-driven alterations. We are using a Whitley i2 Instrument Workstation to study the signalling and metabolic pathways that may preserve breast cancer cell viability under hypoxia. In addition to this, we have a general interest in identifying what signalling and metabolic pathways are associated with the acquisition of microsatellite (MSI) and chromosomal instability (CIN) under oxygen deprivation conditions. Finally, we are studying the role of hypoxia in cell invasion and metastasis, oncogene-induced senescence and resistance to current treatment options.

George Poulogiannis
Team Leader
Division of Cancer Biology
Institute of Cancer Research, London

i2 Workstation Testimonial Video

In this video – the latest in our series of customer testimonials – Dr Ayse Latif, Faculty of Medical and Human Sciences, University of Manchester, discusses her current research of exploiting tumour cell energy metabolism in order to improve gynaecological cancer treatments.

Dr Latif uses both a H35 HEPA Hypoxystation and an i2 Instrument Workstation. Whilst cells are initially incubated in the H35, they are then moved into the i2 via the Whitley Transfer Tunnel without exposure to ambient conditions, where they can be studied with a Seahorse XF Analyzer.

The i2 was designed to meet the exact requirements  of Seahorse Bioscience, meaning that the combination of this workstation and a Seahorse XF Analyzer permits simultaneous, real-time analysis of mitochondrial respiration and glycolysis in mammalian cells under precisely controlled conditions.

sitting-on-beach-using-ipad_web

Did You Know: Whitley Workstations are Ethernet-enabled for Remote Access?

Have you ever wanted to check the status of your Whitley Workstation when you were away from the lab? Well now you can. Most of our new workstations are Ethernet-enabled so you can dial in and access the touch-screen control panel from the other side of the building, or even from the other side of the world. This allows you to check the current operating conditions of the workstation and, if activated, view any of the graphical screens. If necessary, you can then make changes to the parameters, i.e. temperature, humidity, O2 and CO2 levels from wherever, whenever!

Read more

Whitley H35 Hypoxystation

Biological Containment

Looking for extra biological containment from your workstation?

If you need additional biological containment within your HEPA filtered Workstation, there is now an option to reduce biological discharge from the chamber via the gas outlets. The Whitley Enhanced Biological Containment System involves fitting HEPA filters to the two gas outlets on a Whitley HEPA Workstation.

Laboratory tests were conducted on an H35 Hypoxystation to deliberately challenge the exhaust filters with high concentrations of aerosolized bacterial cells and quantify their discharge through the exhaust valve filters.

The results of this experiment demonstrated that the system achieved a substantial reduction in bacterial loading (better than 99.999%) within the discharged gas. A paper detailing the full methodology and results of this study can be found here.

Whitley H35 Hypoxystation

Did You Know: about HEPA Filtration?

HEPA, or High Efficiency Particulate Air, is a filtration system which ensures the utmost product protection for your samples.

Unlike competitor systems that draw atmosphere out of the workstation before filtering and reintroducing it, Don Whitley Scientific has created a unique design, which is housed completely inside the workstation. Housing the HEPA system inside the chamber prevents the build-up of moisture which renders HEPA filters ineffective, and cleans the internal atmosphere quickly.

Using innovative circulation technology, all the atmosphere in the chamber passes through the filter 900 times every hour. This quickly creates and maintains an environment exceeding the level of atmospheric cleanliness stipulated by ISO 14644, referenced in the Cell Tissue Culture Directive as the recommended standard for the handling of mammalian cells. We achieve levels of atmospheric cleanliness exceeding the requirements of Class 3 of this standard, which to the best of our knowledge is better than any other manufacturer.

There are currently three Whitley Workstations available with HEPA filtration:

A35 HEPA Anaerobic Workstation
H35 HEPA Hypoxystation
H45 HEPA Hypoxystation.

Proof of the atmospheric cleanliness achieved during the rigorous tests conducted on every HEPA workstation before dispatch is available on request. Please contact us for more information.

This video demonstrates the effectiveness of our system.

Whitley i2 Instrument Workstation

NEW Whitley i2 Instrument Workstation

The Whitley i2 Instrument Workstation has been developed in response to a rising number of enquiries from scientists who desire to use Seahorse Extracellular Flux (XF) Analyzers* in hypoxic conditions and were dissatisfied with the solutions available. Rather than adapting an existing chamber for this purpose, Don Whitley Scientific developed a new workstation specifically to suit the precise requirements of Seahorse Bioscience.

Extensive research and careful consideration has gone into the development of the i2, which has resulted in a modular system that includes a number of novel and unique features. This workstation can be used as a stand-alone unit or connected to a Whitley Hypoxystation via the new Whitley Transfer Tunnel, enabling preparation of cell lines under hypoxic conditions and their transfer directly into the i2 without exposure to air.

Another unique feature is the integral incubator, which enables you to precondition cellware and incubate plates and media at 37°C under the same atmospheric conditions as the XF Analyzer.

The i2 maintains an internal temperature no higher than 28°C, excludes carbon dioxide, and provides precise oxygen control.

This workstation, the latest in a long line of well-regarded modified atmosphere workstations from Don Whitley Scientific, is equipped with a generous working area in which to conduct preparatory work and is supplied complete with removable front, bespoke trolley, internal mains sockets, and a wireless footswitch to control the patented oval ports.

Whitley i2 Instrument Workstation connected to a H35 HEPA Hypoxystation

Whitley i2 Instrument Workstation connected to a H35 HEPA Hypoxystation

The 12 litre airlock, with a cycle time of just 60 seconds, accommodates up to 44 × 96 well plates or 84 × T25 tissue culture flasks plus numerous other flasks, pipettes and laboratory consumables.

The Whitley i2 Instrument Workstation is the first in a new range of larger Whitley Workstations developed to allow customers to use a variety of items of specialised equipment under modified atmospheric conditions.

The combination of a Whitley i2 Instrument Workstation and Seahorse XF Analyzer permits simultaneous, real-time analysis of mitochondrial respiration and glycolysis in mammalian cells under precisely controlled hypoxic conditions.

If your research requires unsurpassed accuracy, reproducibility and reliability, then insist on a Whitley Workstation.

 

 

* Not presently suitable for the recently announced Seahorse Bioscience XF-p