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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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Interpretation of xCELLigence biosensor Cell Index curves: determination of assay parameters influencing experimental design. After obtaining a standard Cell Index adhesion curve for a particular cell type, it can be used to interpret specific behaviours of the cells relevant to the experimental design. Using the HMEC-1 endothelial cells as an example, the initial period of the Cell Index curve reveals the initial phase of cell adhesion and spreading (0–8 h), followed by a plateau phase prior to a gradual period of proliferation. The arrows represent potential time points for drug treatment; note that the behaviour or response of the cells may vary at each. The curve represents the mean Cell Index value from >4 wells ± SD.
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biosensors-05-00199-f002: Interpretation of xCELLigence biosensor Cell Index curves: determination of assay parameters influencing experimental design. After obtaining a standard Cell Index adhesion curve for a particular cell type, it can be used to interpret specific behaviours of the cells relevant to the experimental design. Using the HMEC-1 endothelial cells as an example, the initial period of the Cell Index curve reveals the initial phase of cell adhesion and spreading (0–8 h), followed by a plateau phase prior to a gradual period of proliferation. The arrows represent potential time points for drug treatment; note that the behaviour or response of the cells may vary at each. The curve represents the mean Cell Index value from >4 wells ± SD.

Mentions: Interpretation of xCELLigence Cell Index curves: The strength of cell adhesion is represented by the software as the Cell Index (unit-less measurement). As cells adhere to the E-plates, the Cell Index value increases from zero, and this will usually be evident within the first 10–15 min of seeding (see Figure 1). Cells that are very strongly adherent will reach a maximum Cell Index of ~10–15. In our experience, this is the upper range for Cell Index values. Non-adherent cells will not affect the Cell Index values greatly (0–1), whereas a Cell Index of 1–4 is weak, with 5–10 being moderate to strong. As adhesion decreases, there will be a concordant decease in the Cell Index. Figure 2 emphasizes different phases of adhesion, which occur for vascular endothelial cells (HMEC-1 cell line). These cells are strongly adherent and also proliferative until they form a monolayer. The initial increase (0–4 h) in the Cell Index is associated with the attachment and adhesion of the cells (schematics in Figure 1b and Figure 2), followed by spreading and a brief plateau phase. This is a proliferative cell line, and the progressive linear increase in the Cell Index post 24 h after seeding is consistent with the initiation of the proliferation of these cells. In theory, this temporal profile should only be observed with proliferating cells (lines), whereas non proliferating cells will remain constant until they become nutrient deprived and compromised.


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)

Interpretation of xCELLigence biosensor Cell Index curves: determination of assay parameters influencing experimental design. After obtaining a standard Cell Index adhesion curve for a particular cell type, it can be used to interpret specific behaviours of the cells relevant to the experimental design. Using the HMEC-1 endothelial cells as an example, the initial period of the Cell Index curve reveals the initial phase of cell adhesion and spreading (0–8 h), followed by a plateau phase prior to a gradual period of proliferation. The arrows represent potential time points for drug treatment; note that the behaviour or response of the cells may vary at each. The curve represents 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-f002: Interpretation of xCELLigence biosensor Cell Index curves: determination of assay parameters influencing experimental design. After obtaining a standard Cell Index adhesion curve for a particular cell type, it can be used to interpret specific behaviours of the cells relevant to the experimental design. Using the HMEC-1 endothelial cells as an example, the initial period of the Cell Index curve reveals the initial phase of cell adhesion and spreading (0–8 h), followed by a plateau phase prior to a gradual period of proliferation. The arrows represent potential time points for drug treatment; note that the behaviour or response of the cells may vary at each. The curve represents the mean Cell Index value from >4 wells ± SD.
Mentions: Interpretation of xCELLigence Cell Index curves: The strength of cell adhesion is represented by the software as the Cell Index (unit-less measurement). As cells adhere to the E-plates, the Cell Index value increases from zero, and this will usually be evident within the first 10–15 min of seeding (see Figure 1). Cells that are very strongly adherent will reach a maximum Cell Index of ~10–15. In our experience, this is the upper range for Cell Index values. Non-adherent cells will not affect the Cell Index values greatly (0–1), whereas a Cell Index of 1–4 is weak, with 5–10 being moderate to strong. As adhesion decreases, there will be a concordant decease in the Cell Index. Figure 2 emphasizes different phases of adhesion, which occur for vascular endothelial cells (HMEC-1 cell line). These cells are strongly adherent and also proliferative until they form a monolayer. The initial increase (0–4 h) in the Cell Index is associated with the attachment and adhesion of the cells (schematics in Figure 1b and Figure 2), followed by spreading and a brief plateau phase. This is a proliferative cell line, and the progressive linear increase in the Cell Index post 24 h after seeding is consistent with the initiation of the proliferation of these cells. In theory, this temporal profile should only be observed with proliferating cells (lines), whereas non proliferating cells will remain constant until they become nutrient deprived and compromised.

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