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Ready-2-Go

Cell Health Assay Services

Assessments of cell health are often limited to classification of cells as ‘live’ or dead’, but cell health is much more nuanced. Multiplexed imaging of cells can provide a more realistic view, but can be difficult to perform. Our R2G Cell Health Assay Service takes a different approach to provide you with better insights into cell health.

Ready-2-Go Assay Services

Microglia Phagocytosis

Key Features & Benefits

  • Control fibrosis induction in 2D model. Cell culture protocols limit stress-fiber formation.

  • Pathologically relevant readout. ɑ-SMA in stress fibers are measured.

  • High-content analysis. Cell morphologies and other phenotypes are measured in addition to ɑ-SMA.

  • Uses a physiologically relevant model. Primary normal human lung fibroblasts (NHLF) cells.

  • R2G assay services are a starting point. Transition to more complex, bespoke assay services with the same service provider.

Key Features & Benefits

  • Physiologically relevant cell models. Wild-type and TREM2-mutant, human iPSC-derived microglia.
  • Greater assurance of data quality. Optimized culture conditions with reference compounds enhance assay reproducibility and minimize noise.

Assays Designed with Your Goals in Mind

Microglia perform a variety of functions essential to establishing and maintaining neuronal health. These cells survey and secrete inflammatory molecules, prune synapses, and are responsible for clearing pathogens, dead cells, and pathological proteins from the extracellular space through phagocytosis. As such, these cells are strongly implicated in the onset or progression of neurodegenerative diseases.

In Alzheimer’s disease (AD), it is hypothesized that M2-like microglia may be beneficial, providing surveillance and phagocytosis of extracellular amyloid and supporting the regeneration and repair of neurons. However, chronic activation may be detrimental; M1-like microglia secrete factors that can damage neurons.

The balance between healthy and disease states are influenced by microglial activities. The modulation of this activity is a potentially powerful strategy to developing therapeutics for neurodegenerative diseases.

The Challenge

To date, assays developed for microglial research have most commonly been conducted using immortalized and murine cell lines, and results from these experiments may not faithfully mirror human biology.

The complex biology and nuanced behavior of microglia, especially in the context of neurodegenerative disease, are still largely unknown, and shortcomings of these models that use microglial substitutes cannot be identified until late in the drug-development process.

However, primary, human microglia are expensive and difficult to access, especially in adequate quantity to screen candidate therapeutics. Challenging culture conditions and patient-to-patient variability further emphasize the need for a reliable, robust, biologically relevant, in vitro system.

Our Solution

Our Ready-2-Go (R2G) Microglia Phagocytosis Assay Service measures the uptake of a fluorescent substrate by iPSC-derived human microglia. iPSC-derived cells afford reproducible, higher-throughput assays. This assay measures the phagocytosis of bioparticles in real-time through live-cell imaging. Reference compounds that activate and inhibit phagocytosis in this system are run in parallel with your test articles.

How It Works

process-timeline

Cells are seeded into 384-well plates. Reference compounds are incubated with the cells overnight, followed by the addition of pHrodo S. aureus bioparticles that fluoresce when phagocytosed into a cell. Fluorescent measurements of the pHrodo intensity and images are taken every 30 minutes for 24 hours. The images and graph below show the phagocytosis of pHrodo bioparticles by wild-type human iPSC-derived microglia treated with reference compounds.

microgliaphago-rep-images-v2
microgliaphago-graph

Assay Service Details

  Ready-2-Go Microglia Phagocytosis Bespoke Assay Services  
Cell Type iCell Microglia (Fujifilm Cellular Dynamics, Inc.): Normal, TREM2 HO, or TREM2 HZ If you would like to expand the service offering beyond R2G shown on the left, please contact us.  
Markers pHrodo bacteria bioparticle, Hoechst (nuclei)  
Dosing 6 doses of your test article  
Positive Control* IL-10  
Negative Controls* Cytochalasin D, IFN-gamma  
Time Points Every 30 min. over 24 hours  
Assay Readouts pHrodo intensity over time  

*Vehicle and untreated controls also included.

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Mission

Mission

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Vision

Vision

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Manifesto

Manifesto

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OUR BUSINESS PARTNERS

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OUR OFFICE

Headquarters, Munich
Pelikanstrasse 2
80333, Munich, DE
POWER TIMELINE

OUR COMPANY HISTORY

  • 2015 - COMPANY START

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  • 2016 - FIRST MILESTONE

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  • 2018 - SECOND MILESTONE

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  • 2020 - TODAY

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