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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

Optimisation of matrix coating for best growth conditions. There are a multitude of factors that can influence the growth and viability of cells. This figure highlights the influence of collagen (red curve) on the adhesion of human brain cerebral microvascular cell (hCMVECs) endothelial cells. Collagen is a major component of the basal lamina of the blood brain barrier structure. Here, the endothelial cells achieve a higher level of adhesion faster than those not grown on a matrix. Mean Cell Index value from >4 wells ± SD.
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biosensors-05-00199-f004: Optimisation of matrix coating for best growth conditions. There are a multitude of factors that can influence the growth and viability of cells. This figure highlights the influence of collagen (red curve) on the adhesion of human brain cerebral microvascular cell (hCMVECs) endothelial cells. Collagen is a major component of the basal lamina of the blood brain barrier structure. Here, the endothelial cells achieve a higher level of adhesion faster than those not grown on a matrix. Mean Cell Index value from >4 wells ± SD.

Mentions: Optimising growth conditions using extracellular matrix coatings: Often, the conditions that produce the correct cellular phenotype or growth conditions for routine cell culture of your cells will produce the best result on xCELLigence technology, too. Figure 4 highlights the difference in the Cell Index for hCMVECs (brain endothelial cells) grown on collagen in comparison to cells grown without matrix. Collagen is an important component of the basement membrane of the blood brain barrier (BBB). It is therefore logical that collagen will improve the adhesion and growth of the brain endothelial cells. The xCELLigence biosensor demonstrates this very easily and can be used to screen which matrix coatings enhance or improve cell growth or viability. Recommendation: the SP system (Single 96-well xCELLigence Plate) provides a good option for conducting the optimisation steps in one or two experiments and can be done using a single plate. Where substantially different curves are obtained on different coatings, consideration should be given as to whether the coating affects the aspects of signaling, growth or cell behaviour relevant to the experimental outcome.


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)

Optimisation of matrix coating for best growth conditions. There are a multitude of factors that can influence the growth and viability of cells. This figure highlights the influence of collagen (red curve) on the adhesion of human brain cerebral microvascular cell (hCMVECs) endothelial cells. Collagen is a major component of the basal lamina of the blood brain barrier structure. Here, the endothelial cells achieve a higher level of adhesion faster than those not grown on a matrix. 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-f004: Optimisation of matrix coating for best growth conditions. There are a multitude of factors that can influence the growth and viability of cells. This figure highlights the influence of collagen (red curve) on the adhesion of human brain cerebral microvascular cell (hCMVECs) endothelial cells. Collagen is a major component of the basal lamina of the blood brain barrier structure. Here, the endothelial cells achieve a higher level of adhesion faster than those not grown on a matrix. Mean Cell Index value from >4 wells ± SD.
Mentions: Optimising growth conditions using extracellular matrix coatings: Often, the conditions that produce the correct cellular phenotype or growth conditions for routine cell culture of your cells will produce the best result on xCELLigence technology, too. Figure 4 highlights the difference in the Cell Index for hCMVECs (brain endothelial cells) grown on collagen in comparison to cells grown without matrix. Collagen is an important component of the basement membrane of the blood brain barrier (BBB). It is therefore logical that collagen will improve the adhesion and growth of the brain endothelial cells. The xCELLigence biosensor demonstrates this very easily and can be used to screen which matrix coatings enhance or improve cell growth or viability. Recommendation: the SP system (Single 96-well xCELLigence Plate) provides a good option for conducting the optimisation steps in one or two experiments and can be done using a single plate. Where substantially different curves are obtained on different coatings, consideration should be given as to whether the coating affects the aspects of signaling, growth or cell behaviour relevant to the experimental outcome.

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