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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

Measurement of rapid or transient cellular responses using xCELLigence. The usefulness of xCELLigence for monitoring acute transient responses is exemplified with the response of microvascular endothelial cells to sphingosine-1-phosphate (S1P). The response to S1P is immediate and transient, and xCELLigence reveals the precise timing and magnitude of the response. S1P was added at several concentrations (addition indicated by black arrow, 1 µM to 1 nM), which reveals a concentration-dependent response. The Normalised Cell Index plot is also used in conjunction with the Cell Index plots to determine the extent (%) of the response. Curves represent the Mean Cell Index value from >4 wells ± SD.
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biosensors-05-00199-f006: Measurement of rapid or transient cellular responses using xCELLigence. The usefulness of xCELLigence for monitoring acute transient responses is exemplified with the response of microvascular endothelial cells to sphingosine-1-phosphate (S1P). The response to S1P is immediate and transient, and xCELLigence reveals the precise timing and magnitude of the response. S1P was added at several concentrations (addition indicated by black arrow, 1 µM to 1 nM), which reveals a concentration-dependent response. The Normalised Cell Index plot is also used in conjunction with the Cell Index plots to determine the extent (%) of the response. Curves represent the Mean Cell Index value from >4 wells ± SD.

Mentions: Identification of acute drug responses: Figure 6 shows the response profile of endothelial cells (HMEC-1 cells) to sphingosine-1-phosphate (S1P; 1 nM to 1 µM). The longitudinal data (three days) in Figure 6a reveal the rapid and transient reduction in adhesion induced by S1P, which occurs immediately following S1P addition (black arrow). Figure 6b focuses on the period immediately after S1P addition to highlight how fast the response occurs. Figure 6c shows the Normalised Cell Index at the time of S1P addition. It reveals that the approximate reduction in adhesion is 10%–20% within 5 min of addition and maximal (~30%) within 10 min. These data show: (I) that the endothelial cells respond to S1P across a range of concentrations; and (II) that the response is immediate and transient. The profile in (a) also demonstrate that the S1P does not induce compromise or death of the cells, which would be shown by a progressive sustained loss in the Cell Index [22]. Although xCELLigence does not reveal the biology of the response, it does reveal that there is a response and most importantly when the response occurs. We are certain that without the real-time biosensor detection, the acute immediate nature of the S1P response would have been missed. This is very useful for future experimental design to identify how S1P regulates endothelial function.


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)

Measurement of rapid or transient cellular responses using xCELLigence. The usefulness of xCELLigence for monitoring acute transient responses is exemplified with the response of microvascular endothelial cells to sphingosine-1-phosphate (S1P). The response to S1P is immediate and transient, and xCELLigence reveals the precise timing and magnitude of the response. S1P was added at several concentrations (addition indicated by black arrow, 1 µM to 1 nM), which reveals a concentration-dependent response. The Normalised Cell Index plot is also used in conjunction with the Cell Index plots to determine the extent (%) of the response. Curves represent the Mean Cell Index value from >4 wells ± SD.
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f006: Measurement of rapid or transient cellular responses using xCELLigence. The usefulness of xCELLigence for monitoring acute transient responses is exemplified with the response of microvascular endothelial cells to sphingosine-1-phosphate (S1P). The response to S1P is immediate and transient, and xCELLigence reveals the precise timing and magnitude of the response. S1P was added at several concentrations (addition indicated by black arrow, 1 µM to 1 nM), which reveals a concentration-dependent response. The Normalised Cell Index plot is also used in conjunction with the Cell Index plots to determine the extent (%) of the response. Curves represent the Mean Cell Index value from >4 wells ± SD.
Mentions: Identification of acute drug responses: Figure 6 shows the response profile of endothelial cells (HMEC-1 cells) to sphingosine-1-phosphate (S1P; 1 nM to 1 µM). The longitudinal data (three days) in Figure 6a reveal the rapid and transient reduction in adhesion induced by S1P, which occurs immediately following S1P addition (black arrow). Figure 6b focuses on the period immediately after S1P addition to highlight how fast the response occurs. Figure 6c shows the Normalised Cell Index at the time of S1P addition. It reveals that the approximate reduction in adhesion is 10%–20% within 5 min of addition and maximal (~30%) within 10 min. These data show: (I) that the endothelial cells respond to S1P across a range of concentrations; and (II) that the response is immediate and transient. The profile in (a) also demonstrate that the S1P does not induce compromise or death of the cells, which would be shown by a progressive sustained loss in the Cell Index [22]. Although xCELLigence does not reveal the biology of the response, it does reveal that there is a response and most importantly when the response occurs. We are certain that without the real-time biosensor detection, the acute immediate nature of the S1P response would have been missed. This is very useful for future experimental design to identify how S1P regulates endothelial function.

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