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Application of xCELLigence RTCA Biosensor Technology for Revealing the Profile and Window of Drug Responsiveness in Real Time.

Kho D, MacDonald C, Johnson R, Unsworth CP, O'Carroll SJ, du Mez E, Angel CE, Graham ES - Biosensors (Basel) (2015)

Bottom Line: In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness.The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay.In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, University of Auckland, Auckland 1023, New Zealand. s.ocarroll@auckland.ac.nz.

ABSTRACT
The xCELLigence technology is a real-time cellular biosensor, which measures the net adhesion of cells to high-density gold electrode arrays printed on custom-designed E-plates. The strength of cellular adhesion is influenced by a myriad of factors that include cell type, cell viability, growth, migration, spreading and proliferation. We therefore hypothesised that xCELLigence biosensor technology would provide a valuable platform for the measurement of drug responses in a multitude of different experimental, clinical or pharmacological contexts. In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness. The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay. In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

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Related in: MedlinePlus

xCELLigence provides quality control and assay variation. The Cell Index curves in (a) show the normal variation that we observe across an E96 well plate, which is usually around 10%–15%. However, xCELLigence can also reveal poor seeding practice (b), which is indicative of such a substantial spread in the Cell Index curves. (c) An example where too few cells were seeded into the E-plate. These exemplify useful teaching and quality control aspects of the technology.
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biosensors-05-00199-f011: xCELLigence provides quality control and assay variation. The Cell Index curves in (a) show the normal variation that we observe across an E96 well plate, which is usually around 10%–15%. However, xCELLigence can also reveal poor seeding practice (b), which is indicative of such a substantial spread in the Cell Index curves. (c) An example where too few cells were seeded into the E-plate. These exemplify useful teaching and quality control aspects of the technology.

Mentions: Revealing good cell culture practice: The real-time measurement of net adhesion in every well provides a valuable quality control measure for cell culture technique. In Figure A2, three very different plots are shown; Figure A2a is an example of the normal variation to expect from good cell culture practice; Figure A2b inconsistent seeding density due to poor pipetting technique; and Figure A2c poor technique in terms of being able to count cells. Each of these plots show the variation in seeding across E96 well plates with each curve representing a separate well. We have noted that large/dense cells, such as astrocytes, can sediment during seeding, which can result in substantial variation in seeding density. xCELLigence reveals this easily and therefore provides an internal quality control. We typically observe 10%–15% variation in the Cell Index across all wells of an E96 well plate, as shown in Figure A2a. However, on occasions and in particular with new practitioners of cell culture, a poor seeding technique Figure A2b or counting technique can be revealed using xCELLigence. This can be an extremely valuable teaching tool for new students. Recommendation: if the level of variation shown in Figure A2b,c is observed with an xCELLigence assay, it is likely to be occurring in other aspects of the cell culture, too!


Application of xCELLigence RTCA Biosensor Technology for Revealing the Profile and Window of Drug Responsiveness in Real Time.

Kho D, MacDonald C, Johnson R, Unsworth CP, O'Carroll SJ, du Mez E, Angel CE, Graham ES - Biosensors (Basel) (2015)

xCELLigence provides quality control and assay variation. The Cell Index curves in (a) show the normal variation that we observe across an E96 well plate, which is usually around 10%–15%. However, xCELLigence can also reveal poor seeding practice (b), which is indicative of such a substantial spread in the Cell Index curves. (c) An example where too few cells were seeded into the E-plate. These exemplify useful teaching and quality control aspects of the technology.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4493546&req=5

biosensors-05-00199-f011: xCELLigence provides quality control and assay variation. The Cell Index curves in (a) show the normal variation that we observe across an E96 well plate, which is usually around 10%–15%. However, xCELLigence can also reveal poor seeding practice (b), which is indicative of such a substantial spread in the Cell Index curves. (c) An example where too few cells were seeded into the E-plate. These exemplify useful teaching and quality control aspects of the technology.
Mentions: Revealing good cell culture practice: The real-time measurement of net adhesion in every well provides a valuable quality control measure for cell culture technique. In Figure A2, three very different plots are shown; Figure A2a is an example of the normal variation to expect from good cell culture practice; Figure A2b inconsistent seeding density due to poor pipetting technique; and Figure A2c poor technique in terms of being able to count cells. Each of these plots show the variation in seeding across E96 well plates with each curve representing a separate well. We have noted that large/dense cells, such as astrocytes, can sediment during seeding, which can result in substantial variation in seeding density. xCELLigence reveals this easily and therefore provides an internal quality control. We typically observe 10%–15% variation in the Cell Index across all wells of an E96 well plate, as shown in Figure A2a. However, on occasions and in particular with new practitioners of cell culture, a poor seeding technique Figure A2b or counting technique can be revealed using xCELLigence. This can be an extremely valuable teaching tool for new students. Recommendation: if the level of variation shown in Figure A2b,c is observed with an xCELLigence assay, it is likely to be occurring in other aspects of the cell culture, too!

Bottom Line: In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness.The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay.In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

View Article: PubMed Central - PubMed

Affiliation: Department of Pharmacology, University of Auckland, Auckland 1023, New Zealand. s.ocarroll@auckland.ac.nz.

ABSTRACT
The xCELLigence technology is a real-time cellular biosensor, which measures the net adhesion of cells to high-density gold electrode arrays printed on custom-designed E-plates. The strength of cellular adhesion is influenced by a myriad of factors that include cell type, cell viability, growth, migration, spreading and proliferation. We therefore hypothesised that xCELLigence biosensor technology would provide a valuable platform for the measurement of drug responses in a multitude of different experimental, clinical or pharmacological contexts. In this manuscript, we demonstrate how xCELLigence technology has been invaluable in the identification of (1) not only if cells respond to a particular drug, but (2) the window of drug responsiveness. The latter aspect is often left to educated guess work in classical end-point assays, whereas biosensor technology reveals the temporal profile of the response in real time, which enables both acute responses and longer term responses to be profiled within the same assay. In our experience, the xCELLigence biosensor technology is suitable for highly targeted drug assessment and also low to medium throughput drug screening, which produces high content temporal data in real time.

Show MeSH
Related in: MedlinePlus