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

Direct comparison of growth characteristics of NT2-astrocytes from different cultures. Here, xCELLigence was used to assess the consistency and growth characteristics of NT2-derived astrocytes produced from different differentiations (and by different students). This example shows that the astrocytes in Differentiation C had a very low Cell Index, which is inconsistent with the strong level of adhesion typical of these cells (Differentiations A and B).
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f005: Direct comparison of growth characteristics of NT2-astrocytes from different cultures. Here, xCELLigence was used to assess the consistency and growth characteristics of NT2-derived astrocytes produced from different differentiations (and by different students). This example shows that the astrocytes in Differentiation C had a very low Cell Index, which is inconsistent with the strong level of adhesion typical of these cells (Differentiations A and B).

Mentions: Comparison of variation were made between differentiations or donors (in our research, we use a range of cells derived from cell lines [5,20,21] (e.g., HEKs, HMEC-1 and hCMVECs), differentiated precursors (NT2 astrocytes [7,22,23] and neurons [24]) and from primary cells from blood [25,26] and brain [27]). The simple set-up and real-time readout of xCELLigence provides a technology to directly compare any variation in the global temporal growth characteristics of cells from different batches of differentiated cells (e.g., NT2) or from different donors. Figure 5a shows the Cell Index growth curves from three independent NT2 astrocyte differentiations. These cells are produced following an expensive 10-week differentiation protocol. Typically, we assess the success of the differentiation using GFAP/vimentin/βIII-tubulin expression to indicate the purity and yield of astrocytes. The data shown in Figure 5 clearly suggest that the growth of the astrocytes from Differentiation C (represented by the blue Cell Index curve) was substantially different from that of A and B (red and green curves). We have assessed more than 30 different astrocyte differentiations using xCELLigence, and they typically produce a Cell Index >7 (strong). The weak adhesion from Culture C suggested that the yield or health of the astrocytes was poor and therefore not suitable for further assays. Recommendation: in addition to comparison of differentiations (yield, quality and consistency), the long-term temporal capacity of xCELLigence is ideal for comparing the consistency of cell lines over time, especially those that may have been passaged to high numbers or genetically modified. In addition, it has applications for primary cells from different donors or the comparison between different disease states.


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)

Direct comparison of growth characteristics of NT2-astrocytes from different cultures. Here, xCELLigence was used to assess the consistency and growth characteristics of NT2-derived astrocytes produced from different differentiations (and by different students). This example shows that the astrocytes in Differentiation C had a very low Cell Index, which is inconsistent with the strong level of adhesion typical of these cells (Differentiations A and B).
© Copyright Policy
Related In: Results  -  Collection

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

biosensors-05-00199-f005: Direct comparison of growth characteristics of NT2-astrocytes from different cultures. Here, xCELLigence was used to assess the consistency and growth characteristics of NT2-derived astrocytes produced from different differentiations (and by different students). This example shows that the astrocytes in Differentiation C had a very low Cell Index, which is inconsistent with the strong level of adhesion typical of these cells (Differentiations A and B).
Mentions: Comparison of variation were made between differentiations or donors (in our research, we use a range of cells derived from cell lines [5,20,21] (e.g., HEKs, HMEC-1 and hCMVECs), differentiated precursors (NT2 astrocytes [7,22,23] and neurons [24]) and from primary cells from blood [25,26] and brain [27]). The simple set-up and real-time readout of xCELLigence provides a technology to directly compare any variation in the global temporal growth characteristics of cells from different batches of differentiated cells (e.g., NT2) or from different donors. Figure 5a shows the Cell Index growth curves from three independent NT2 astrocyte differentiations. These cells are produced following an expensive 10-week differentiation protocol. Typically, we assess the success of the differentiation using GFAP/vimentin/βIII-tubulin expression to indicate the purity and yield of astrocytes. The data shown in Figure 5 clearly suggest that the growth of the astrocytes from Differentiation C (represented by the blue Cell Index curve) was substantially different from that of A and B (red and green curves). We have assessed more than 30 different astrocyte differentiations using xCELLigence, and they typically produce a Cell Index >7 (strong). The weak adhesion from Culture C suggested that the yield or health of the astrocytes was poor and therefore not suitable for further assays. Recommendation: in addition to comparison of differentiations (yield, quality and consistency), the long-term temporal capacity of xCELLigence is ideal for comparing the consistency of cell lines over time, especially those that may have been passaged to high numbers or genetically modified. In addition, it has applications for primary cells from different donors or the comparison between different disease states.

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