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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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xCELLigence technology; impedance recording and cell adhesion. Summary of xCELLigence system technology and impedance, which shows: (a) the high density gold array on the E-plates; and (b) the initial phases of seeding and adhesion, which is recorded by the biosensor and represented as the Cell Index (adhesion).
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biosensors-05-00199-f001: xCELLigence technology; impedance recording and cell adhesion. Summary of xCELLigence system technology and impedance, which shows: (a) the high density gold array on the E-plates; and (b) the initial phases of seeding and adhesion, which is recorded by the biosensor and represented as the Cell Index (adhesion).

Mentions: The E16 or E96 xCELLigence plates were prepared by addition of complete media (50 µL) to every well. After equilibration to 37 °C, plates were inserted into the xCELLigence station, and the base-line impedance was measured to ensure that all wells and connections were working within acceptable limits. The software automatically informs the researcher if any connection problems arise. Following harvesting and counting, cells were diluted to the correct seeding density and added to the wells in 50 µL (see the schematic in Figure 1). We have found that this volume provides less variation in seeding density, especially for larger/dense cells, like astrocytes. We would recommend that researchers should seed their cells using their standard seeding protocols and assess how much seeding variation exists.


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 technology; impedance recording and cell adhesion. Summary of xCELLigence system technology and impedance, which shows: (a) the high density gold array on the E-plates; and (b) the initial phases of seeding and adhesion, which is recorded by the biosensor and represented as the Cell Index (adhesion).
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f001: xCELLigence technology; impedance recording and cell adhesion. Summary of xCELLigence system technology and impedance, which shows: (a) the high density gold array on the E-plates; and (b) the initial phases of seeding and adhesion, which is recorded by the biosensor and represented as the Cell Index (adhesion).
Mentions: The E16 or E96 xCELLigence plates were prepared by addition of complete media (50 µL) to every well. After equilibration to 37 °C, plates were inserted into the xCELLigence station, and the base-line impedance was measured to ensure that all wells and connections were working within acceptable limits. The software automatically informs the researcher if any connection problems arise. Following harvesting and counting, cells were diluted to the correct seeding density and added to the wells in 50 µL (see the schematic in Figure 1). We have found that this volume provides less variation in seeding density, especially for larger/dense cells, like astrocytes. We would recommend that researchers should seed their cells using their standard seeding protocols and assess how much seeding variation exists.

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