Live Cell Insights Publications Newsletter
Read our summaries of new and notable research articles using the IncuCyte® Live Cell Analysis System, including hand-picked publications for applications and research areas like neuroinflammation, cell motility, development, and more!
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IncuCyte® has reached 2,700 cited publications spanning a wide array of research areas and applications. We’ve experienced over 50% growth in publications in just the last year! Search our publications list to see what exciting research is being published using the IncuCyte® Live-Cell Analysis System.
Effective, well-tolerated treatments to help curb the immune-mediated damage to the central nervous system during multiple sclerosis (MS) have remained elusive. The dysregulation of glutamate and activation of T-cells are known to be involved in disease pathogenesis, spurring a number of pharmacological attempts to treat disease effects with limited success.
In this study by Hollinger and colleagues at Johns Hopkins, the drug JHU-083 (a liphophilic glutamine antagonist) was tested in an attempt to overcome peripheral toxicity encountered in the use of DON (6-diazo-5-oxo-L-norleucine) drug candidates and their limited penetration into the brain. JHU-083, a liphophilic prodrug candidate of DON, was designed to be inert in the circulation to reduce toxicity, but upon penetration into the brain, becomes cleaved and is activated. The antiproliferative effects on T-cell proliferation of the drug were examined in vitro, along with additional in vivo physical and cognitive assessment in a mouse model of MS, experimental autoimmune encephalomyelitis (EAE).
Key findings include:
JHU-083 inhibited the in vivo proliferation of T cells and decreased their activation, as determined by flow cytometry. There was no effect on bone marrow derived dendritic cells.
To confirm these findings, and to determine if the anti-proliferative effects of JHU-083 were due to drug cytotoxicity, IncuCyte® Live-Cell Imaging and Analysis was used to measure proliferation and viability of CD4+ T cells isolated from the naive mouse spleens. The activation of CD4+ T cells was followed using the IncuCyte reagents Cytotox Green (for cell death) and NucLight Rapid Red (for cell number). IncuCyte analysis revealed that all doses of the drug caused reduced CD4+ proliferation and reduced cell number, even the lower doses, which was important for reduced toxicity. Further, IncuCyte analysis also revealed that the drug was not cytotoxic as compared to DMSO control.
In vivo administration of JH-083 reduced the severity of EAE symptoms in two testing conditions: prevention and treatment. Most interestingly, the cognitive impairment associated with this EAE murine model was reversed, as assessed by Barnes maze testing.
JHU-083 shows promise as a novel potential therapeutic for the treatment of physical and cognitive symptoms of MS, which may also have important implications for other neuroinflammatory-related conditions.
Read the full paper in Neurology, Neuroimmunology & Neuroinflammation, August 2019.
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IncuCyte® Cytotoxicity Assays
The IncuCyte® Cytotoxicity Assay uses the IncuCyte® Cytotox Reagents to make real time measurements of cell death based on cell membrane integrity, all within your incubator.
Actin is a critical component of the cytoskeleton, the organization and dynamic assembly of which are involved in changes in cellular shape, motility, and processes such as gene transcription. Post-translational modifications of the actin protein, such and N-terminal (Nt)-acetylation, can affect cellular functions but this process has thus far been uncharacterized.
In this study by Drazic, Aksnes, and Marie, et al. of the Arnesen lab at the University of Bergen, the authors report on the exciting discovery of actin’s N-terminal acetyltransferase: NAA80. Digging deeper, this work investigates the various activities of Nt-acetylation that affect cytoskeletal morphology and cell hypermotility, emphasizing how this important actin modification impacts both functional and cellular responses, opening an avenue to manipulate this process through gene targeting of NAA80.
Key findings include:
Read the full paper in PNAS, April 2018.
Watch the supplemental video of wound closure from this paper.
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IncuCyte® Scratch Wound Cell Migration and Invasion System
An integrated solution enables real-time visualization and assessment of cell morphology in scratch wound assays (both label free and fluorescently labeled) up to six 96-well plates at once – all inside your tissue culture incubator.
Cdc42 is a GTPase that regulates signaling for a variety of cell functions in vitro, including changes in cellular morphology, cell polarity, the migration of cells, and cell cycle. This is accomplished via activation of some families of kinases, which may influence the actin organization and cellular migration. Though these effects have been studied in vitro, the in vivo effects have not been well characterized.
In this international, collaborative study by Lavina et al., the authors examined the regulatory role of small GTPase cell division cycle 42 (Cdc42) on endothelial cell functions, angiogenesis, and vascular morphogenesis in a mouse retinal model. The authors combined the power of conditional, mosaic gene ablation and computational modeling to identify the cellular functions that were dependent on Cdc42, and which were related to vascular morphogenesis.
Key findings include:
Read the full paper in Development, July 2018.
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IncuCyte® Image Lock Plates
The IncuCyte® ImageLock Plates when used in conjunction with IncuCyte® software, allow high-quality image registration for time-lapse imaging. The technology enables a user to revisit and image the same cells at multiple time points and is especially useful for multi-day, long-term live-cell imaging applications.
The importance of the interactions between RNA-binding proteins and miRNAs in the control of gene expression is becoming more apparent, and such interactions may be crucial for determining cell fate, neurogenesis, brain development, and neurological disease pathology.
In this multi-institutional study by Velasco et al., the authors built upon their previous finding that tumor suppressor miRNAs regulate Musashi1 (Msi1), an RNA binding protein and stem cell marker, which also regulates cell fate either towards self-renewal or cellular differentiation. In particular, the miRNA miR-137 can upregulate Msi1 to promote differentiation, thereby preventing the development of glioblastoma. In this follow-up investigation, the authors attempted to further characterize this relationship, proposing an antagonistic model, whereby by Msi1 and miR-137 act to regulate via activation or repression, influencing the determination of cell fate.
Key findings include:
Read the full paper in RNA, April 2019.
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IncuCyte® Neuronal Cell Health and Morphology Assays
Neuronal Cell Health and Morphology Assays reveal both significant and subtle dynamic changes in cells over an extended period of time using fully automated image acquisition and analysis.
Lung cancer continues to be the leading cause of cancer deaths, partially due to late–stage diagnosis and resulting delay in treatment. This necessitates the continued, urgent search for more effective therapeutics. Of particular interest for drug targeting are the endogenous signal transducer and activator of transcription (STAT) proteins, which are involved in various aspects of the immune response and may be downregulated during tumor development.
In this multidisciplinary study from the University of Michigan, Speth et al. creatively approached this challenge by developing synthetic liposomes for the delivery of an endogenous STAT inhibitor suppressor of cytokine signaling (SOCS3). This agent was utilized both in vitro, and in an in vivo in a lung cancer xenograft mouse model. The authors attempted to harness the power of an endogenous antitumor mechanism, performing a therapeutic rescue investigation based on their previous discovery that alveolar macrophages (AM) curb inflammatory responses in lung epithelial cells (ECs).
Key findings include:
Read the full paper in JCI Insight, October 2019.
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IncuCyte® Apoptosis Assays
The mix-and-read IncuCyte® Caspase-3/7 and Annexin V Reagents let you measure multiple apoptotic pathways simultaneously and in real time...
Many cancer treatments, such as chemotherapy and radiation, inflict DNA damage on tumors. The inability of a tumor to repair this DNA damage could theoretically increase treatment efficacy, but this requires further investigation and characterization.
In this study by Essers et al. from the Netherlands Cancer Institute and VUmc Cancer Center, the authors utilized machine learning for the development of gene expression models. They identified functional defects in gene repair pathways in head and neck squamous cell carcinoma (HNSCC), with subsequent validation and impact on patient prognosis. This paper shows the power of an efficient screening mechanism for DNA repair defect to gain important insights for the HNSCC patient response to chemo-radiotherapy and outcome assessment.
Key findings include:
Read the full paper in Cancer Research, September 2019.
FEATURED PRODUCT
IncuCyte® Scratch Wound Cell Migration and Invasion System
An integrated solution enables real-time visualization and assessment of cell morphology in scratch wound assays (both label free and fluorescently labeled) up to six 96-well plates at once – all inside your tissue culture incubator.