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xCELLigence system for real-time label-free monitoring of growth and viability of cell lines from hematological malignancies.

Martinez-Serra J, Gutierrez A, Muñoz-Capó S, Navarro-Palou M, Ros T, Amat JC, Lopez B, Marcus TF, Fueyo L, Suquia AG, Gines J, Rubio F, Ramos R, Besalduch J - Onco Targets Ther (2014)

Bottom Line: With a fibronectin substrate, nonadherent cells deposited in a monolayer configuration, and consequently, the cell growth and viability were robustly monitored.In order to validate this technology, the data obtained through real-time cell analysis was compared with that obtained from using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.This provides an excellent label-free tool for the screening of drug efficacy in nonadherent cells and discriminates optimal time points for further molecular analysis of cellular events associated with treatments, reducing both time and costs.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, University Hospital Son Espases, Palma de Mallorca, Balearic Islands, Spain.

ABSTRACT
The xCELLigence system is a new technological approach that allows the real-time cell analysis of adherent tumor cells. To date, xCELLigence has not been able to monitor the growth or cytotoxicity of nonadherent cells derived from hematological malignancies. The basis of its technology relies on the use of culture plates with gold microelectrodes located in their base. We have adapted the methodology described by others to xCELLigence, based on the pre-coating of the cell culture surface with specific substrates, some of which are known to facilitate cell adhesion in the extracellular matrix. Pre-coating of the culture plates with fibronectin, compared to laminin, collagen, or gelatin, significantly induced the adhesion of most of the leukemia/lymphoma cells assayed (Jurkat, L1236, KMH2, and K562). With a fibronectin substrate, nonadherent cells deposited in a monolayer configuration, and consequently, the cell growth and viability were robustly monitored. We further demonstrate the feasibility of xCELLigence for the real-time monitoring of the cytotoxic properties of several antineoplastic agents. In order to validate this technology, the data obtained through real-time cell analysis was compared with that obtained from using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. This provides an excellent label-free tool for the screening of drug efficacy in nonadherent cells and discriminates optimal time points for further molecular analysis of cellular events associated with treatments, reducing both time and costs.

No MeSH data available.


Related in: MedlinePlus

Fibronectin pre-coating significantly increases Jurkat, K562, and KMH2 cell attachment to the bottom of the culture well.Notes: (A) Fibronectin induced a rapid B-CI increase (<2 hours) in Jurkat cells. The maximum B-CI value within 2 hours is included for each substrate. The B-CI is presented as means ± SD from at least two (up to four) independent wells (calculated by xCELLigence). (B) Fibronectin unlike other substrates also increased the B-CI for KMH2 and K562. For U937 cells, not even the fibronectin succeeded in attaching leukemia cells to the bottom of the well.Abbreviations: B-CI, baseline cell index; SD, standard deviation.
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f1-ott-7-985: Fibronectin pre-coating significantly increases Jurkat, K562, and KMH2 cell attachment to the bottom of the culture well.Notes: (A) Fibronectin induced a rapid B-CI increase (<2 hours) in Jurkat cells. The maximum B-CI value within 2 hours is included for each substrate. The B-CI is presented as means ± SD from at least two (up to four) independent wells (calculated by xCELLigence). (B) Fibronectin unlike other substrates also increased the B-CI for KMH2 and K562. For U937 cells, not even the fibronectin succeeded in attaching leukemia cells to the bottom of the well.Abbreviations: B-CI, baseline cell index; SD, standard deviation.

Mentions: We first pre-coated the E-Plate surfaces (1 hour) with 6 μg of fibronectin, collagen, laminin, or 0.2% volume per weight of gelatin. We then seeded Jurkat cells on the E-Plates. In the absence of a coating substrate, after 2 hours, the B-CI was almost 0 (mean B-CI: 0.1169±0.0658). On the other hand, fibronectin, compared to the other substrates, allowed a significant and rapid increase of the B-CI (mean B-CI: 0.7106±0.0635) (cell attaching stage), suggesting a more efficient adhesion induced by the presence of this coating substrate (Figure 1A). After 24 hours, the RTCA values for Jurkat cells reached a B-CI of 0.8849±0.0311 for fibronectin, 0.1997±0.0378 for collagen, 0.16720±0221 for gelatin, and B-CI: 0.0570±0.0273 for laminin (Figure 1A). We next incubated KMH2 cells, K562, and U937 in the presence of all these substrates. In Figure 1B, we show that under the same experimental conditions, almost all the leukemia/lymphoma cell lines tested showed a significant increased B-CI only in the presence of fibronectin. On the other hand, for U937 cells, even fibronectin failed to attach leukemia cells at the bottom of the well.


xCELLigence system for real-time label-free monitoring of growth and viability of cell lines from hematological malignancies.

Martinez-Serra J, Gutierrez A, Muñoz-Capó S, Navarro-Palou M, Ros T, Amat JC, Lopez B, Marcus TF, Fueyo L, Suquia AG, Gines J, Rubio F, Ramos R, Besalduch J - Onco Targets Ther (2014)

Fibronectin pre-coating significantly increases Jurkat, K562, and KMH2 cell attachment to the bottom of the culture well.Notes: (A) Fibronectin induced a rapid B-CI increase (<2 hours) in Jurkat cells. The maximum B-CI value within 2 hours is included for each substrate. The B-CI is presented as means ± SD from at least two (up to four) independent wells (calculated by xCELLigence). (B) Fibronectin unlike other substrates also increased the B-CI for KMH2 and K562. For U937 cells, not even the fibronectin succeeded in attaching leukemia cells to the bottom of the well.Abbreviations: B-CI, baseline cell index; SD, standard deviation.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4061162&req=5

f1-ott-7-985: Fibronectin pre-coating significantly increases Jurkat, K562, and KMH2 cell attachment to the bottom of the culture well.Notes: (A) Fibronectin induced a rapid B-CI increase (<2 hours) in Jurkat cells. The maximum B-CI value within 2 hours is included for each substrate. The B-CI is presented as means ± SD from at least two (up to four) independent wells (calculated by xCELLigence). (B) Fibronectin unlike other substrates also increased the B-CI for KMH2 and K562. For U937 cells, not even the fibronectin succeeded in attaching leukemia cells to the bottom of the well.Abbreviations: B-CI, baseline cell index; SD, standard deviation.
Mentions: We first pre-coated the E-Plate surfaces (1 hour) with 6 μg of fibronectin, collagen, laminin, or 0.2% volume per weight of gelatin. We then seeded Jurkat cells on the E-Plates. In the absence of a coating substrate, after 2 hours, the B-CI was almost 0 (mean B-CI: 0.1169±0.0658). On the other hand, fibronectin, compared to the other substrates, allowed a significant and rapid increase of the B-CI (mean B-CI: 0.7106±0.0635) (cell attaching stage), suggesting a more efficient adhesion induced by the presence of this coating substrate (Figure 1A). After 24 hours, the RTCA values for Jurkat cells reached a B-CI of 0.8849±0.0311 for fibronectin, 0.1997±0.0378 for collagen, 0.16720±0221 for gelatin, and B-CI: 0.0570±0.0273 for laminin (Figure 1A). We next incubated KMH2 cells, K562, and U937 in the presence of all these substrates. In Figure 1B, we show that under the same experimental conditions, almost all the leukemia/lymphoma cell lines tested showed a significant increased B-CI only in the presence of fibronectin. On the other hand, for U937 cells, even fibronectin failed to attach leukemia cells at the bottom of the well.

Bottom Line: With a fibronectin substrate, nonadherent cells deposited in a monolayer configuration, and consequently, the cell growth and viability were robustly monitored.In order to validate this technology, the data obtained through real-time cell analysis was compared with that obtained from using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method.This provides an excellent label-free tool for the screening of drug efficacy in nonadherent cells and discriminates optimal time points for further molecular analysis of cellular events associated with treatments, reducing both time and costs.

View Article: PubMed Central - PubMed

Affiliation: Department of Hematology, University Hospital Son Espases, Palma de Mallorca, Balearic Islands, Spain.

ABSTRACT
The xCELLigence system is a new technological approach that allows the real-time cell analysis of adherent tumor cells. To date, xCELLigence has not been able to monitor the growth or cytotoxicity of nonadherent cells derived from hematological malignancies. The basis of its technology relies on the use of culture plates with gold microelectrodes located in their base. We have adapted the methodology described by others to xCELLigence, based on the pre-coating of the cell culture surface with specific substrates, some of which are known to facilitate cell adhesion in the extracellular matrix. Pre-coating of the culture plates with fibronectin, compared to laminin, collagen, or gelatin, significantly induced the adhesion of most of the leukemia/lymphoma cells assayed (Jurkat, L1236, KMH2, and K562). With a fibronectin substrate, nonadherent cells deposited in a monolayer configuration, and consequently, the cell growth and viability were robustly monitored. We further demonstrate the feasibility of xCELLigence for the real-time monitoring of the cytotoxic properties of several antineoplastic agents. In order to validate this technology, the data obtained through real-time cell analysis was compared with that obtained from using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. This provides an excellent label-free tool for the screening of drug efficacy in nonadherent cells and discriminates optimal time points for further molecular analysis of cellular events associated with treatments, reducing both time and costs.

No MeSH data available.


Related in: MedlinePlus