Applications


Cancer Immunotherapy OverviewWhat is cancer immunotherapy? Cancer immunotherapy consists of multiple approaches that focus on harnessing and enhancing the innate powers of the immune system to fight cancer. While traditional small molecule chemotherapy continues to play a critical role in cancer treatment, immunotherapy is rapidly gaining traction; in 2014 immunotherapies constituted ~50% of the overall oncology pharmacopeia. Cancer immunotherapies can be divided ... Read More
Cancer Immunotherapy: Antibody-Dependent Cell-Mediated Cytotoxicity (ADCC)What is ADCC? Though the innate and adaptive branches of the immune system are typically described as being distinct and separate from one another, they often work in concert to afford protection and combat disease. Upon encountering a pathogen, cells of the innate immune system typically release cytokines that cross-talk with components of the adaptive immune system, causing them to expand and become ... Read More
Cancer Immunotherapy: Bispecific T Cell Engagers (BiTEs) and Bispecific AntibodiesWhat are Bispecific Antibodies/Bispecific T Cell Engagers (BiTEs)? The therapeutic efficacy of the antibody-dependent cell-mediated cytotoxicity (ADCC) technique is mitigated by the fact that not all immune cells express the CD16 antibody receptor. In particular, cytotoxic and helper T lymphocytes don’t express CD16 and therefore aren’t recruited to antibody-coated cells. In order to circumvent this constraint and mobilize the full capacity ... Read More
Cancer Immunotherapy: Genetically Engineered T Cell-Mediated Cell KillingGenetically Engineered T Cells T cells can be genetically engineered to express a modified T cell receptor (TCR; specific for a tumor antigen) or a chimeric antigen receptor (CAR; composed of an intracellular signaling domain that is linked to an extracellular domain derived from a tumor-specific antibody). Avoiding the immune tolerance issues associated with non-autologous therapies, and producing T cells ... Read More
Cancer Immunotherapy: Macrophage-Mediated PhagocytosisTumor Associated Macrophages Macrophages are important effector cells of innate immunity. Depending on the tissue microenvironment, tumor-associated macrophages (TAM) can differentiate into either cytotoxic (M1) or tumor-promoting (M2) states. While cytotoxic M1 macrophages are typically induced by IFN-γ alone or in concert with microbial products, tumor promoting M2 macrophages are induced by IL-4 or IL-13, IL-10, IL-21, TGFβ, immune complexes, ... Read More
Cancer Immunotherapy: NK Cell-Mediated CytolysisWhat are NK Cells? Natural killer (NK) cells are a type of cytotoxic lymphocyte that play a critical role in the innate immune system, primarily by recognizing and destroying virus-infected cells. NK cells express a number of activation and inhibitory receptors that work in concert to distinguish infected or diseased cells from normal cells. In close proximity to a target cell slated for ... Read More
Cancer Immunotherapy: T Cell-Mediated CytolysisWhat are T Cells? By seeking out and destroying infected cells directly, the CD8+ subgroup of T cells play a critical role in the adaptive immune response. Every CD8+ T cell clone expresses a unique variant of a specialized receptor, the T cell receptor (TCR), that can recognize and bind to a specific antigenic peptide presented by MHC class I ... Read More
Cardiosafety: Drug-Induced ArrythmiaBetween 1990 and 2006 one third of all safety-related drug withdrawals were due to cardiotoxicity, and cardiac liability continues to be a major hurdle in drug development.  Within the pharmaceutical industry, academic institutions, and regulatory agencies there are ongoing efforts to develop higher throughput and more predictive assays that can be used earlier in the drug discovery/development pipeline to minimize ... Read More
Cell AdhesionQuantitative Assessment of Extracellular Matrix Effect on A549 Attachment and Spreading The Cell Index increases proportionately as the coating concentration of collagen IV increases. (Data and figures adapted from ACEA Biosciences, unpublished data). Mammary Stem-Like Cells Have Increased Tubulin-Dependent Initial Reattachment From Suspension (A) Stem-like HMLE cells attach at signifi cantly faster rates than non-stem-like HMLE cells as determined by ... Read More
Cell Barrier FunctionComparison of Conventional Methods with the Dynamic Impedance-Based Method for Monitoring Ethanol-Induced Epithelial Barrier Dysfunction (A) The reversible barrier dysfunction induced by 7.5% ethanol was assessed by measuring phenol red permeability (control: untreated Caco-2 cell monolayer; EtOH: Caco-2 exposed in ethanol for 3 h; EtOH removal: Caco-2 exposed in ethanol for 3 h followed by replacing ethanol with fresh medium ... Read More
Cell Invasion and Migration OverviewCell migration and invasion are critical to many facets of biology, including such diverse processes as wound healing and the intravasation/extravasation of cancer cells during metastasis. A major hurdle to studying migration and invasion has been the lack of techniques that are quantitative, reproducible, and efficient. Scratch assays/wound healing assays require repetitive visual evaluation of cells and provide subjective data ... Read More
Cell Migration/Invasion: Chemotactic MigrationSee the Cell Migration/Invasion Overview for background information. The examples below demonstrate the use of ACEA’s cell invasion and migration plate (CIM-Plate®) and the xCELLigence® RTCA DP system to study chemotactic migration, with specific focus on inflammation and cancer metastasis. APPLICATION HIGHLIGHT: Real-Time Chemotaxis of Macrophages Macrophage recruitment to sites of tissue injury, inflammation, or infection is essential for mounting ... Read More
Cell Migration/Invasion: TransmigrationSee the Cell Migration/Invasion Overview for background information. The example below demonstrates the use of ACEA’s cell invasion and migration plate (CIM-Plate®) and the xCELLigence® RTCA DP system to study transmigration, with specific focus on cancer spheroid invasion. APPLICATION HIGHLIGHT: Studying Ovarian Cancer Spheroid Invasion of Mesothelial Cells Once ovarian cancer cells shed into the peritoneal fluid, they aggregate and ... Read More
Cell-Cell Interations: Co-CultureGrowth Factor Induced Proliferation of T47D Cells Monitoring of T47D cellular response to H295R estrogen signaling. Estrogen and progesterone receptor positive T47D breast cancer cells were grown on xCELLigence E-plates. After a coculture insert containing H295R adrenal corticocarcinoma cells, known to produce estrogen and progesterone, was placed on the E-plate. The response of the T47D cells to the secreted hormones ... Read More
Cell-Response ProfilingUsing the xCELLigence System for Target Identification and Validation (A) HeLa cells were transfected with various concentrations of KIF11 (the gene encoding Eg5) siRNA. A time and concentration-dependent antimitotic time-dependent cell response profiles (TCRPs) was observed with higher siRNA concentrations producing more pronounced CI changes. The kinetics of the CI profiles is very similar between various concentrations of siRNAs: CI ... Read More
Cytotoxicity OverviewCytotoxicity is the general quality of being toxic to cells, and can be caused by chemical stimuli, exposure to other cells (NK or T cells for example), or physical/environmental conditions (radiation exposure, temperature or pressure extremes, etc.).  Cytotoxic treatment can result in one of three potential cellular fates.  Whereas necrotic cell death involves the rapid loss of membrane integrity and ... Read More
Cytotoxicity: Compound-Mediated CytotoxicityKinetic Response Profiles Reflect Different Cytotoxic Mechanisms HepG2 cells were treated with various cytotoxic compounds and monitored in real time for cytotoxic response kinetics. Representative compounds targeting tubulin (paclitaxel), actin (cytochalasin D), DNA (doxorubicin), mitochondria (rotenone), calcium homeostasis (thapsigargin), and protein transport (brefeldin A) generated distinct response profiles. (Data and figure adapated from ACEA Biosciences, Inc. unpublished data). Real Time ... Read More
Cytotoxicity: NanotoxicityNanomaterials are organic or inorganic particles whose size is less than 100 nanometers in at least one dimension.  Though nanomaterials hold tremendous promise for myriad applications, their impact on living cells in the short and long term are incompletely understood.  Numerous studies have suggested that nanomaterials, even when made of inert materials, can still display toxicity because of their physical ... Read More
Functional Analysis of Genes & Proteins: siRNAUsing the xCELLigence System for Target Identification and Validation (A) HeLa cells were transfected with various concentrations of KIF11 (the gene encoding Eg5) siRNA.  A time and concentration-dependent antimitotic time-dependent cell response profiles (TCRPs) was observed with higher siRNA concentrations producing more pronounced CI changes. The kinetics of the CI profiles is very similar between various concentrations of siRNAs: CI ... Read More
Immune Cell ActivationReal-Time Monitoring of Mast Cell Degranulation in RBL-2H3 Mast Cell Line (A) Real-time acquisition of impedance-based measurements captures immediate kinetic profi le changes of RBL-2H3 when sensitized with IgE and subsequently stimulated with DNP-BSA. (B) Rhodamine-phalloidin staining of sensitized and activated RBL-2H3 cells performed in parallel to the impedance assay indicates that stimulated cells undergo cytoskeleton rearrangements which correlate with the ... Read More
In Vitro Hypoxia StudiesHypoxia, where the body or localized regions of the body are deprived of adequate oxygen supply, occurs during numerous acute and chronic disease states.  Examples include reduced blood flow during heart attack or stroke, and the oxygen-restricted microenvironment within a tumor.  Though cells can be studied in vitro under similar oxygen-poor conditions using a hypoxia chamber glove box, the inefficiencies ... Read More
Parasitic Worm Motility and ViabilityDespite the prevalence and expense of parasitic worm (helminth) infection worldwide, the helminth-specific pharmacopeia is extremely limited.  This is historically due, in part, to the paucity of objective high-throughput drug screening methods amenable to the life cycle of these parasites.  Though hampered by observer subjectivity and low throughput capacity, for many years the gold standard assay for anthelmintic drug screening ... Read More
Phenotypic ScreeningPhenotypic Screening: Probing Efficacy and Mechanism of Action Simultaneously Because the ultimate effect of drugs are the net result of on-target and less well understood off-target interactions, the assays traditionally employed in drug discovery and development usually suffer from at least one limitation.  Due to their readout being so narrow, target-focused biochemical assays have a higher probability of failing to ... Read More
Quality Control of CellsThough a great deal of attention is typically paid to optimizing the conditions and reagents used in cell-based assays, quality control of the cells themselves is frequently overlooked.  Human errors such as mislabeling or cross contaminating cell lines can lead to researchers working with cells of unintended identity. Moreover, even in the absence of errors/contamination, the inherent genetic instability of ... Read More
Receptor Signaling OverviewCritical to all forms of life is the ability to temporally regulate cellular processes based on environment/circumstance.  Whereas the downstream regulatory processes can involve biochemical modification (phosphorylation, allosteric inhibition, etc.) and/or changes to transcription/translation profiles, the initial “sensing” event occurs predominantly through membrane receptor signaling (Figure 1).   Figure 1.  Membrane receptor signaling.  Extracellular stimuli are initially “sensed” predominantly through ... Read More
Receptor Signaling: GPCR-mediated signalingGPCR-Mediated Signaling G protein-coupled receptors (GPCRs) are the most abundant family of cell surface receptors and represent the largest class of therapeutic targets.  Responding to a wide range of stimuli, GPCRs elicit diverse signaling events ranging from phosphorylation cascades and transcription regulation to ion channel activity and secondary messenger production.  Either directly or indirectly these signaling events usually lead to ... Read More
Receptor Signaling: Nuclear Hormone-Mediated SignalingEstrogen Specific Response Monitored on xCELLigence System The estrogen receptor agonist 17β-estradiol (E2) induced a unique kinetic response profile with delayed cell index growth in the human breast cancer cell line T-47D. Such response was abolished by the estrogen receptor antagonist ICI182780 (A). The progesterone antagonist mifepristone cannot abolish E2 response (B), but rather had a synergistic effect with E2 ... Read More
Receptor Signaling: RTK-Mediated SignalingEndogenous Receptor Tyrosine Kinase Short Term Response Cells expressing recombinant PDGFRb were seeded on an E-plate 96 at 20,000 cells per well and after overnight growth the medium was replaced with serum free medium (not shown). (A) After 2 hours cells were treated with increasing doses of the PDGF nhibitor Imatinib, incubated for one hour, then stimulated with 10 ng/ml ... Read More
Stem Cell and Cell DifferentiationReal-Time Monitoring and Image Analysis of Human Skin-Derived Precursor (SKPs) Cell Differentiation Differentiation of human skin-derived precursors (SKPs) into smooth muscle cells (SMCSs) and undifferentiated SKPs (C) with or without the application of transforming growth factor (TGF)-β1 (A) or TGF-β31 (B) and subsequently fixed and stained to display a-smooth actin, calponin, and SM22a. Real-time analysis correlates the increase in Cell Index with ... Read More
Virus-Mediated Cytopathic Effect (CPE)Virus infection of a host cell typically includes the selective suppression of host cell functions and redirection of resources towards viral replication and assembly, ultimately leading to host cell lysis and dissemination of new virus.  While host cell rounding, detachment from the plate surface and/or lysis are readily detected by real-time impedance monitoring, more subtle changes in host cell morphology ... Read More
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