Limits...
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.

Show MeSH

Related in: MedlinePlus

xCELLigence reveals the differential temporal responsiveness of skin endothelial cells to various TLR ligands across a six-day time course. CL075 (5 μg/mL) induces an immediate and sustained increase in endothelial adhesion, whereas the R837 (5 μg/mL) effects are not obvious until at least 48 h after drug addition. The cells did not appear to respond to R848 (10 μg/mL). The black arrow indicates the time of TLR agonist addition, and the acute response period, highlighted by the red box, is shown in the lower panel. The lower panel reveals clearly the immediacy of the CL075 response. Each curve represents the mean of four wells ± SD. Note that the lower panel is normalised to the time of drug addition.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4493546&req=5

biosensors-05-00199-f008: xCELLigence reveals the differential temporal responsiveness of skin endothelial cells to various TLR ligands across a six-day time course. CL075 (5 μg/mL) induces an immediate and sustained increase in endothelial adhesion, whereas the R837 (5 μg/mL) effects are not obvious until at least 48 h after drug addition. The cells did not appear to respond to R848 (10 μg/mL). The black arrow indicates the time of TLR agonist addition, and the acute response period, highlighted by the red box, is shown in the lower panel. The lower panel reveals clearly the immediacy of the CL075 response. Each curve represents the mean of four wells ± SD. Note that the lower panel is normalised to the time of drug addition.

Mentions: The data in Figure 8 are a prime example of how xCELLigence can reveal drug responses that occur in very different time frames. In this study, a specific panel of TLR ligands was assessed using xCELLigence for those that regulated skin endothelial cells. We had no prior knowledge of whether the skin endothelial cells would respond, as the majority of TLR work is conducted with immune cells from blood. This select panel comprised CL075, R848 (Gardiquimod) and R837 (Imiquimod). CL075 is a thiazoloquinolone derivative that stimulates TLR8 and, at a higher concentration, can activate TLR7 in human peripheral blood mononuclear cells and in these cells is known to activate NF-κB, resulting in the production of pro-inflammatory mediators, including TNF-α and IL-12 [28]. The imidazoquinoline compound R848 (Resiquimod) is also an agonist for TLR7 and TLR8. R837 is an FDA-approved drug (active ingredient in Aldara and Zyclara) and prescription secondary medication for various skin cancers and also indicated for warts. Pharmacologically, R837 targets TLR7 and is thought to mediate its clinical effects through the activation of skin antigen-presenting cells. The goal of our experiments was to assess whether skin endothelial cells (HMEC-1 cells) responded to any of the TLR ligands in the panel and to ascertain the window of drug responsiveness (if any).


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 reveals the differential temporal responsiveness of skin endothelial cells to various TLR ligands across a six-day time course. CL075 (5 μg/mL) induces an immediate and sustained increase in endothelial adhesion, whereas the R837 (5 μg/mL) effects are not obvious until at least 48 h after drug addition. The cells did not appear to respond to R848 (10 μg/mL). The black arrow indicates the time of TLR agonist addition, and the acute response period, highlighted by the red box, is shown in the lower panel. The lower panel reveals clearly the immediacy of the CL075 response. Each curve represents the mean of four wells ± SD. Note that the lower panel is normalised to the time of drug addition.
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f008: xCELLigence reveals the differential temporal responsiveness of skin endothelial cells to various TLR ligands across a six-day time course. CL075 (5 μg/mL) induces an immediate and sustained increase in endothelial adhesion, whereas the R837 (5 μg/mL) effects are not obvious until at least 48 h after drug addition. The cells did not appear to respond to R848 (10 μg/mL). The black arrow indicates the time of TLR agonist addition, and the acute response period, highlighted by the red box, is shown in the lower panel. The lower panel reveals clearly the immediacy of the CL075 response. Each curve represents the mean of four wells ± SD. Note that the lower panel is normalised to the time of drug addition.
Mentions: The data in Figure 8 are a prime example of how xCELLigence can reveal drug responses that occur in very different time frames. In this study, a specific panel of TLR ligands was assessed using xCELLigence for those that regulated skin endothelial cells. We had no prior knowledge of whether the skin endothelial cells would respond, as the majority of TLR work is conducted with immune cells from blood. This select panel comprised CL075, R848 (Gardiquimod) and R837 (Imiquimod). CL075 is a thiazoloquinolone derivative that stimulates TLR8 and, at a higher concentration, can activate TLR7 in human peripheral blood mononuclear cells and in these cells is known to activate NF-κB, resulting in the production of pro-inflammatory mediators, including TNF-α and IL-12 [28]. The imidazoquinoline compound R848 (Resiquimod) is also an agonist for TLR7 and TLR8. R837 is an FDA-approved drug (active ingredient in Aldara and Zyclara) and prescription secondary medication for various skin cancers and also indicated for warts. Pharmacologically, R837 targets TLR7 and is thought to mediate its clinical effects through the activation of skin antigen-presenting cells. The goal of our experiments was to assess whether skin endothelial cells (HMEC-1 cells) responded to any of the TLR ligands in the panel and to ascertain the window of drug responsiveness (if any).

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