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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

Use of xCELLigence for measuring drug effects on cell viability, compromise and death. Here, NT2 astrocytes were treated with the pro-inflammatory cytokine IL1β at a range of concentration to assess the acute and long-term effects on the cells. During the initial 24-h period following cytokine treatment, there is a pronounced increase in the Cell Index, which is consistent with inflammatory activation of the cell and increased astrocytic size as a consequence. This is followed by a progressive and continual loss of adhesion. IL1β was added 24 h after seeding, and each curve represents the mean ± SD of four wells. The images in (b) show the viable astrocytes remaining on the E-plate at the end of the experiment. The white dotted circles reveal the position of the non-transparent electrode array. These images were acquired using an inverted microscope. The array can be seen more easily in the bright-field insert. Note that ACEA have developed plates with view strips for imaging of cells during xCELLigence experiments.
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biosensors-05-00199-f007: Use of xCELLigence for measuring drug effects on cell viability, compromise and death. Here, NT2 astrocytes were treated with the pro-inflammatory cytokine IL1β at a range of concentration to assess the acute and long-term effects on the cells. During the initial 24-h period following cytokine treatment, there is a pronounced increase in the Cell Index, which is consistent with inflammatory activation of the cell and increased astrocytic size as a consequence. This is followed by a progressive and continual loss of adhesion. IL1β was added 24 h after seeding, and each curve represents the mean ± SD of four wells. The images in (b) show the viable astrocytes remaining on the E-plate at the end of the experiment. The white dotted circles reveal the position of the non-transparent electrode array. These images were acquired using an inverted microscope. The array can be seen more easily in the bright-field insert. Note that ACEA have developed plates with view strips for imaging of cells during xCELLigence experiments.

Mentions: The data in Figure 7 are unpublished data from an earlier study investigating the influence of various inflammatory cytokines on the function of NT2-derived astrocytes [22,23]. The longitudinal capacity of xCELLigence revealed that several pro-inflammatory cytokines (IL-1β and TNFα) induced compromise of the astrocytes at specific concentrations. This is highlighted in Figure 7a, exemplifying the effect of IL-1β. The initial increase in the Cell Index is consistent with the increased size and activity of the astrocytes during the initial inflammatory phase, which is soon followed by gradual progressive compromise and death. The response was concentration dependent (5 pg/mL to 50 ng/mL). In this study, the xCELLigence profile suggested that compromise was occurring 24–48 h after cytokine addition. This was confirmed using a combination of terminal assays, including cell counts, nuclear fragmentation and cleaved caspase-3 expression [22]. It is important to note that the cytokine-induced compromise of the astrocytes was only realised due to the long-term xCELLigence data. In [22], we utilized the real-time readout of xCELLigence as a precise guidance system to conduct terminal assays to determine the extent of compromise and death with conventional apoptotic assays [22] and cell counts (Figure 7b).


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)

Use of xCELLigence for measuring drug effects on cell viability, compromise and death. Here, NT2 astrocytes were treated with the pro-inflammatory cytokine IL1β at a range of concentration to assess the acute and long-term effects on the cells. During the initial 24-h period following cytokine treatment, there is a pronounced increase in the Cell Index, which is consistent with inflammatory activation of the cell and increased astrocytic size as a consequence. This is followed by a progressive and continual loss of adhesion. IL1β was added 24 h after seeding, and each curve represents the mean ± SD of four wells. The images in (b) show the viable astrocytes remaining on the E-plate at the end of the experiment. The white dotted circles reveal the position of the non-transparent electrode array. These images were acquired using an inverted microscope. The array can be seen more easily in the bright-field insert. Note that ACEA have developed plates with view strips for imaging of cells during xCELLigence experiments.
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f007: Use of xCELLigence for measuring drug effects on cell viability, compromise and death. Here, NT2 astrocytes were treated with the pro-inflammatory cytokine IL1β at a range of concentration to assess the acute and long-term effects on the cells. During the initial 24-h period following cytokine treatment, there is a pronounced increase in the Cell Index, which is consistent with inflammatory activation of the cell and increased astrocytic size as a consequence. This is followed by a progressive and continual loss of adhesion. IL1β was added 24 h after seeding, and each curve represents the mean ± SD of four wells. The images in (b) show the viable astrocytes remaining on the E-plate at the end of the experiment. The white dotted circles reveal the position of the non-transparent electrode array. These images were acquired using an inverted microscope. The array can be seen more easily in the bright-field insert. Note that ACEA have developed plates with view strips for imaging of cells during xCELLigence experiments.
Mentions: The data in Figure 7 are unpublished data from an earlier study investigating the influence of various inflammatory cytokines on the function of NT2-derived astrocytes [22,23]. The longitudinal capacity of xCELLigence revealed that several pro-inflammatory cytokines (IL-1β and TNFα) induced compromise of the astrocytes at specific concentrations. This is highlighted in Figure 7a, exemplifying the effect of IL-1β. The initial increase in the Cell Index is consistent with the increased size and activity of the astrocytes during the initial inflammatory phase, which is soon followed by gradual progressive compromise and death. The response was concentration dependent (5 pg/mL to 50 ng/mL). In this study, the xCELLigence profile suggested that compromise was occurring 24–48 h after cytokine addition. This was confirmed using a combination of terminal assays, including cell counts, nuclear fragmentation and cleaved caspase-3 expression [22]. It is important to note that the cytokine-induced compromise of the astrocytes was only realised due to the long-term xCELLigence data. In [22], we utilized the real-time readout of xCELLigence as a precise guidance system to conduct terminal assays to determine the extent of compromise and death with conventional apoptotic assays [22] and cell counts (Figure 7b).

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