Applications

The facilities from the ImpedanCELL platform enable high-throughput studies in areas as wide-ranging as oncology, neuroscience, immunology, virology, bacteriology or marine biology. At both sites, we have created an "xCELLothèque" in order to respond quickly to the requests we receive answer from the scientific community as well as to develop industrial partnerships. For the Baclesse site, we have about 30 cell models (cell lines and primary cells) whose culture conditions have been defined and validated (number of cells, cell quality index, characteristic impedance profile, response to treatment, etc.). For the LABÉO site, a catalog referencing all the cell types (about 15 to date) and available viruses in different animal species is being set up. The objective is to provide a characteristic impedance profile for each cell/virus system in order to propose models for studying viruses that infect various animal species (target species such as horses, cattle, birds, etc.) in industrial and academic laboratories.

Applications Overview on the xCELLigence systems

The panel of the most developed applications on xCELLigence systems allows the study of the adhesion, the proliferation, the viability, the cytotoxicity, the differentiation as well as the changes of cellular morphology or the functional analysis of receptors (GPCR, RTK) with direct applications in many fields (in particular in oncology, neurology, cardiology, toxicology, immunology, virology, marine biology...). In addition to the measurement of cellular activity in real-time, the CELLAVISTA® microscope allows a daily monitoring of cellular morphology, then offering complementary information, which gives clues about the nature of the observed phenomenon (adhesion, cellular detachment, proliferation, cell death, epithelial-mesenchymal transition, differentiation, etc.).

As illustrated in the figure across the text, the increase of the cellular index in response to the treatment of cells by a cytotoxic agent (carboplatin) can be related to their swelling and to the increase of the adhesion. On the other hand, the measurement of cellular activity in real-time and the additional information provided by the morphological observation of the cells allow to conclude about a differential effect between a molecule 1 (cytotoxic) and a molecule 2 (cytostatic).

Study of the cytotoxicity of different compounds on
an ovarian chemoresistant cancer cell line

Study of the migration and the invasion of HeLa cells

The xCELLigence RTCA DP system model has the capability of performing real-time cell (without the use of labels) invasion/migration assays using CIM-Plate, which is an electronically integrated Boyden chamber. For example, HeLa cells are placed in the upper chamber either directly on top of the microporous membrane containing 8 μm pores (migration assay) or on top of a basement membrane matrix previously deposited on the membrane (invasion assay). Moving towards chemoattractant in the lower chamber, cells pass through the microporous membrane and then adhere to gold impedance microelectrodes. The resultant change in impedance signal correlates with the number of cells attached to these electrodes, enabling easy and fast collection of reproducible data over time ranges spanning from minutes to days.

For example, the ImpedanCELL core facility has been used in the field of oncology to perform a high throughput screening in one hand of a miRNA bank to identify cytotoxic miRNAs and in the other hand of potential inhibiting molecules of the Bcl-2 family anti-apoptotic proteins and in the field of virology to screen a chemical library in order to identify molecules with antiviral properties.

Criblage fonctionnel d’une banque de 1200 miARNs pour identifier des apoptomiRs

Functional Screening of a bank of 1200 miRNAs to identify apoptomiRs.

Identification of the cytotoxic action of miR-491-5p in an ovarian cancer
line (Denoyelle et al., Cell Death Dis, 2014).

Identification of Mcl-1 inhibitors by screening.

Identification of pyridoclax as a potential inhibitor of Mcl-1 in cancer cells addicted to anti-apoptotic Bcl xL and Mcl-1 (Gloagen et al., J Med Chem, 2015).

Screening of antiviral compounds against EHV-1 (Equine Herpes Virus).

Study of the antiviral effect of the acyclovir against EHV-1 in an equine cellular model
(Hue et al., JEVS, 2016, 39: S69-S70).

Among the multiple applications that can be envisaged with the real-time cellular imaging system (IncuCyte S3), the ImpedanCELL platform has been used, for example, to characterize apoptotic cell death in real-time using fluorescent reagents (green caspase-3 kit), to analyze the ability of cancer cells to reinvest an injured space (healing or wound healing) and to follow the appearance of a cytopathic effect of a virus on a cell culture.

Real-time cell death analysis (Caspase-3/7 Green Apoptosis assay)

Study of the ability of cancer cells to reinvest an injured
space (Scratch Wound Migration assay)

Study of the cytopathogenic effect of an equine virus