cat no | io1096 Early Access
GFP ioMicroglia Male
Male human iPSC donor-derived microglia constitutively expressing GFP
GFP ioMicroglia are built from our well-established wild type ioMicroglia Male, deterministically programmed using opti-ox technology and engineered to constitutively express green fluorescent protein (GFP).
Assay ready in just 10 days, these cells offer researchers a fluorescent human microglia model ideal for co-culture with other neural cell types, enabling effortless tracking in multi-cellular systems.
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Confidently investigate your phenotype of interest across multiple clones with our disease model clone panel. Detailed characterisation data (below) and bulk RNA sequencing data (upon request) help you select specific clones if required.
Benchtop benefits
Co-culture compatible
Easily track microglia integration into multi-cell and 3D in vitro models. Ready for co-culture with neurons from day 1.
Functional
Constitutively express GFP throughout the cytosol. Demonstrate key phagocytic and cytokine secretion functions.
Live-cell imaging ready
Study microglia motility and identify different activation states in real-time.
Co-culture protocol
Easy-to-use co-culture protocol for GFP ioMicroglia with ioGlutamatergic Neurons
This co-culture protocol describes a method of co-culturing GFP ioMicroglia with ioGlutamatergic Neurons and associated disease models.
Live-cell imaging in mixed cultures
Live-cell imaging reveals clear visualisation of GFP ioMicroglia when co-cultured with ioGlutamatergic Neurons
ioGlutamatergic Neurons (io1001) were cultured to day 10 post-thaw. GFP ioMicroglia were cultured to day 10 post-thaw and were directly added to day 11 ioGlutamatergic Neurons. The co-cultures were maintained for a further 3 days before live-cell imaging with Leica DMi8. Brightfield and fluorescence images were taken and merged, easily demonstrating distribution of GFP ioMicroglia within the co-culture.
Pro-inflammatory cytokine secretion
GFP ioMicroglia secrete pro-inflammatory cytokines upon activation
GFP ioMicroglia were stimulated at day 10 post-thaw with LPS 100 ng/ml and IFNɣ 20 ng/ml for 24 hours. Supernatants were harvested and analysed using MSD V-plex Proinflammatory Kit. GFP ioMicroglia secrete TNF⍺, IL-6, IL-8, IL-1b, IL-12p70 and IL-10 in response to the inflammatory stimuli. GFP ioMicroglia demonstrate a similar response to ioMicroglia Male (io1021), as expected. Three technical replicates were performed per lot.
View the stimulation for cytokine release protocol used to generate this data.
Phagocytosis functionality
Phagocytosis of E. coli particles by GFP ioMicroglia
(A) Phagocytosis assay using pHrodo™ E. coli BioParticles™ at day 10 post-thaw demonstrates efficient uptake of bacteria particles by GFP ioMicroglia in comparison to ioMicroglia Male (io1021) +/- cytochalasin D control (an inhibitor of actin polymerisation). The graph displays that the proportion of cells phagocytosing E.coli particles over 24 hours for three technical replicates.
(B) The graph displays that the degree of cells phagocytosing E.coli particles over 24 hours. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. Three technical replicates were performed.
GFP ioMicroglia phagocytose Zymosan particles
Representative video shows GFP-labelled ioMicroglia phagocytosing pHrodo Red-labeled Zymosan particles. Upon engulfment, the acidic environment of the phagolysosome causes the particles to fluoresce red, enabling particle visualisation within the cells. Live-cell imaging was conducted at 4-minute intervals over 1.5 hours using the Leica DMi8 microscope.
Highly characterised and defined
Flow cytometry analysis of GFP expression at day 11 and day 21 for GFP ioMicroglia
Flow cytometry analysis demonstrating GFP expression in over 99.5% of cells for GFP ioMicroglia cultured until day 11, with no GFP expression seen in ioMicroglia Male (io1021). At day 21, the percentage of cells expressing GFP and the intensity does not decrease over time, indicating there is no silencing of the reporter gene.
GFP ioMicroglia express key microglia markers
Immunofluorescent staining on day 10 post-revival demonstrates similar homogenous expression of microglia markers P2RY12 and IBA1 and ramified morphology in GFP ioMicroglia compared to ioMicroglia Male (io1021). GFP expression can be visualised throughout the cytosol in every cell for GFP ioMicroglia, but not in the WT control. 10X magnification.
GFP ioMicroglia show ramified morphology by day 10
GFP ioMicroglia mature rapidly, and key ramified morphology can be identified by day 4 and continues through to day 10, similarly to ioMicroglia Male (io1021). Day 1 to 10 post-thawing; 100x magnification.
Ready within days
GFP ioMicroglia Male are delivered in a cryopreserved format and are programmed to rapidly mature upon revival in the recommended media. The protocol for the generation of these cells is a three-phase process: an Induction phase that is carried out at bit.bio, Phase 1: Stabilisation for 24 hours, Phase 2: Maturation for a further 9 days, Phase 3: the Maintenance phase. Cells are ready to use from day 10.
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Human iPSC-derived cells
powered by opti-ox
Consistent. Defined. Scalable.
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ioMicroglia Customer Testimonials
Matteo Zanella, PhD
Associate Research Leader | Charles River