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Roche Diagnostics Deutschland GmbH Sandhofer Str. 116 68309 Mannheim, Deutschland http://www.roche.de/
Ansprechpartner:in Dr. Burkhard Ziebolz +49 8856 604830
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Roche Diagnostics Deutschland GmbH

Label-free Measurement of Natural Killer Cell Cytotoxic Activity in Real-Time with the New xCELLigence System from Roche

(lifePR) (Penzberg, )
Natural killer (NK) cells have been implicated in the recognition and annihilation of tumor cells. As demonstrated in a series of experiments, the real-time impedance-based, label-free xCELLigence System from Roche offers the only assay format so far to directly monitor NK cell mediated cytolysis without labelling the target cells and without using any chemical reporters. In addition, the technology that was invented by ACEA Biosciences, a cooperation partner of Roche in San Diego, USA, allows more accurate assessment of cytolytic activity by dynamically monitoring the entire history of cytolysis.

Natural Killer (NK) cells play a prominent role in various physiological and disease states. The assessment of their cytolytic activity is not only important for monitoring of immunocompetence in cancer, infectious diseases, and autoimmune diseases, but also in determining the proteins which mediate the cytolytic effect.

The standard methods used for measuring NK cell cytolytic activity are radioactive release assays, radio-labelling the target cells with for example 51Cr, and then adding the effector cells, such as NKs. The extent of NK activity towards target cells is assayed by the release of radioactivity by the target cells upon cytolysis. However, these methods are cumbersome not only due to the use of radioactivity but also due to high background because of the tendency of the label to permeate out of the target cells.

Monitoring of NK mediated cytotoxicity by measuring electronic impedance in real-time As shown in a series of experiments, the use of the novel, real time, impedance-based xCELLigence System offers an alternative for label-free in vitro research of NK cell-mediated cytotoxic activity with significant contributions in ease of use and quality of data. The core of the xCELLigence System is the electrical impedance cell sensor arrays integrated into the bottom of microtiter plates which provide continuous, quantitative information about the biological status of attached cells. Any changes to cell number, size, morphology, or attachment quality will be detected in real-time without the use of reporter reagents. The integrated software allows data to be collected and saved automatically as often as every minute over virtually any time period.

Accurate assessment of NK-mediated cytolytic activity

The new impedance-based technology was used by researchers to dynamically and quantitatively monitor NK-mediated cytotoxic activity towards nine different target cell lines, including human cancer cell lines commonly used in the research field. Target cells were seeded in the wells of E-Plate microtiter plates and the impedance-based system dynamically monitored the cell growth every 60 minutes until the cells reached growth phase. The effector murine NK cells were then directly added to the wells at different E/T ratios, and the NK cell-mediated cytolysis was dynamically monitored. In addition, the technoloy was also successfully used in antibody mediated cell cytolysis assay (ADCC) for dynamic and quantitative monitoring of antibody mediated cancer cell killing activity.

In summary, these experiments demonstrate that the xCELLigence System can be used for assessment of NK cell-mediated cytolytic activity in both human and murine NK cell lines. The quantitative, dynamic measurement of NK cell-mediated cytolysis was achieved on the system without any labelling steps or reagents. Moreover, data obtained by the xCELLigence System clearly show that cytolytic activity can reach up to 70 %, which occurs after twelve hours of adding mNK cells. Such cytolytic activity which occurs after the standard incubation time of the assay can be easily missed by existing label-based endpoint assays. Therefore, the xCELLigence System not only offers label-free detection but, more importantly, also allows more accurate assessment of cytolytic activity by dynamically monitoring the entire history of cytolysis.

For more information on the technology, please visit www.xCELLigence.roche.com

About ACEA
ACEA Biosciences has pioneered the development of high-performance microelectronic systems for cell-based assays. ACEA and its partner Roche will market on a world-wide basis ACEA's microplate-based biosensor platform for a variety of applications in drug development, toxicology, cancer research, and medical microbiology and virology. ACEA's products are designed to meet the increasing needs of the life science research and drug discovery markets by providing cost-effective, high content, real time detection systems that simplify complex cell-based assay procedures and increase productivity. With its international reach, ACEA Biosciences works closely with scientists throughout the world. Hangzhou, China is the site of ACEA's world-class manufacturing operations. ACEA Biosciences headquarters is in San Diego, USA.

Roche Diagnostics Deutschland GmbH

Headquartered in Basel, Switzerland, Roche is one of the world's leading research-focused healthcare groups in the fields of pharmaceuticals and diagnostics. As the world's biggest biotech company and an innovator of products and services for the early detection, prevention, diagnosis and treatment of diseases, the Group contributes on a broad range of fronts to improving people's health and quality of life. Roche is the world leader in in-vitro diagnostics and drugs for cancer and transplantation, and is a market leader in virology. It is also active in other major therapeutic areas such as autoimmune diseases, inflammatory and metabolic disorders and diseases of the central nervous system. In 2007 sales by the Pharmaceuticals Division totaled 36.8 billion Swiss francs, and the Diagnostics Division posted sales of 9.3 billion francs. Roche has R&D agreements and strategic alliances with numerous partners, including majority ownership interests in Genentech and Chugai, and invested over 8 billion Swiss francs in R&D in 2007. Worldwide, the Group employs about 80,000 people. Additional information is available on the Internet at www.roche.com.

NIMBLEGEN is a trademark of Roche.

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Für die oben stehenden Stories, das angezeigte Event bzw. das Stellenangebot sowie für das angezeigte Bild- und Tonmaterial ist allein der jeweils angegebene Herausgeber (siehe Firmeninfo bei Klick auf Bild/Titel oder Firmeninfo rechte Spalte) verantwortlich. Dieser ist in der Regel auch Urheber der Texte sowie der angehängten Bild-, Ton- und Informationsmaterialien. Die Nutzung von hier veröffentlichten Informationen zur Eigeninformation und redaktionellen Weiterverarbeitung ist in der Regel kostenfrei. Bitte klären Sie vor einer Weiterverwendung urheberrechtliche Fragen mit dem angegebenen Herausgeber. Bei Veröffentlichung senden Sie bitte ein Belegexemplar an service@lifepr.de.