cat no | io1021
ioMicroglia Male
Male human iPSC donor-derived microglia
ioMicroglia (io1021) are male donor-derived human microglial cells, deterministically programmed using opti-ox technology.
Within 10 days post-revival, ioMicroglia are ready for experimentation, expressing (>90%) key microglia markers, including TMEM119, P2RY12, IBA1, TREM2, CX3CR1, CD11b, CD45, and CD14.
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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
Functional
ioMicroglia display key phagocytic and cytokine secretion functions with lot-to-lot consistency.
Quick
Rapidly maturing male donor-derived human microglia that are ready to use within 10 days post-revival.
Co-culture compatible
Suitable for co-culture studies with neurons at 1 day post-thaw.
Ready within days
opti-ox precision deterministically programmed ioMicroglia from a male donor rapidly form a homogenous microglia population.
Time-lapse video capturing the rapid and homogeneous microglia phenotype acquisition upon thawing of cryopreserved male donor-derived ioMicroglia. 10 day time course.
Highly characterised and defined
Flow cytometry analysis of male donor-derived ioMicroglia shows key phenotypic marker expression
Flow cytometry analysis of day 10 male donor-derived ioMicroglia shows key microglia marker expression of TMEM119, P2RY12, CD14, CD45 and CD11b with a purity of above 95% for CD45, CD11b and CD14, >80% for TMEM119 and CD45, and >70% for TMEM119 and P2RY12.
View the cell detachment protocol used to generate this data.
Male donor-derived ioMicroglia show key microglia marker expression
Immunofluorescent staining of day 10 male donor-derived ioMicroglia shows homogenous expression of P2RY12, IBA1 and TREM2, and a typical ramified morphology. DAPI counterstain (blue). Image taken at 10x magnification.
Male donor-derived ioMicroglia show ramified morphology by day 10
Rapid morphological changes in the cells upon reprogramming, with key ramified morphology first identified by day 4 and continuing through to day 10. Day 1 to 10 post-thawing; 100x magnification; scale bar; 400 µm.
Principal component analysis of bulk RNA sequencing data from male donor-derived ioMicroglia, integrated with sequencing data from Abud et al. (1) shows that these cells cluster closely to primary foetal and adult microglia data sets derived from this publication. Shapes represent the experiment from which data was obtained and colours represent the cell type.
(1) Abud E, et al., Neuron, 2018; 94(2): 278-293
Key functions with consistency
Phagocytosis of E. coli particles by male donor-derived ioMicroglia
Day 10 male donor-derived ioMicroglia from three independent lots were incubated with 1 µg/0.33 cm2 pHrodo RED labelled E. coli particles for 24 hours +/- cytochalasin D control. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. The graph displays the proportion of cells phagocytosing E. coli particles over 24 hours. Three technical replicates were performed per lot.
Degree of phagocytosis of E. coli particles by male donor-derived ioMicroglia
Day 10 male donor-derived ioMicroglia from three independent lots were incubated with 1 µg/0.33 cm2 pHrodo RED labelled E. coli particles for 24 hours +/- cytochalasin D control. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. The graph displays the fluorescence intensity per cell displaying degree of phagocytosis per cell, data from three independent lots. Three technical replicates were performed per lot.
Phagocytosis of Amyloid Beta-42 particles by male donor-derived ioMicroglia
Male donor-derived ioMicroglia show a pro-inflammatory cytokine response to LPS and dexamethasone treatment
(2) Malika Bsibsi, Kimberly Lo, Matteo Zanella, Lieke Geerts, and Stefan Kostense, Charles River Laboratories, Darwinweg 24, 2333 Leiden, The Netherlands.
Male donor-derived ioMicroglia show a pro-inflammatory cytokine response to LPS and Amyloid Beta stimulation
(3) Eve Corrie, Emma V. Jones, Medicines Discovery Catapult, Block 35, Mereside, Alderley Park, Macclesfield, SK10 4ZF. UK.
C5a-mediated chemotaxis
Male donor-derived ioMicroglia display C5a-mediated chemotaxis in a dose-dependent manner
Male donor-derived ioMicroglia were seeded and matured in a plate for 1 week before adding C5a (chemoattractant) and scanning for cell migration. The chemotaxis assay was performed using Clearview plates with the Incucyte® Chemotaxis Analysis Software module. A and B, confluency in the insert (top area) decreased over time as the cells migrated towards C5a, via pores in the insert into a reservoir containing C5a, in a dose-dependent manner. Corresponding confluency in the reservoir (bottom area) increased over time. C and D, Maximal migration of cells is observed at 10 nM with a 5-fold increase in cells present in the reservoir at 60 hours. Explore the full dataset.
This data was generated by Eve Corrie and Emma V. Jones from Medicines Discovery Catapult (3).
(3) Eve Corrie, Emma V. Jones, Medicines Discovery Catapult, Block 35, Mereside, Alderley Park, Macclesfield, SK10 4ZF. UK.
Co-culture compatible
ioGlutamatergic Neurons (io1001) were cultured to day 10 post-thaw. Male donor-derived ioMicroglia (io1021) cultured to either day 1 or day 10 post-thaw were added directly to day 10 ioGlutamatergic Neurons. The co-cultures were maintained for a further 8 days. Imaging was performed in 30-minute intervals. Representative video showing that these cells form a stable co-culture with ioGlutamatergic Neurons.
Key marker expression in male donor-derived ioMicroglia and ioGlutamatergic Neuron co-cultures
Immunofluorescent analysis at day 8 of the co-cultures shows expression of the microglia marker, IBA1 (yellow) and the pan-neuronal marker, MAP2 (red), as expected. Representative images taken at 10x magnification with 100 μm scale bar.
Male donor-derived ioMicroglia retain phagocytic function in co-culture with ioGlutamatergic Neurons
Representative video showing male donor-derived ioMicroglia in co-culture with ioGlutamatergic Neurons selectively phagocytosing pHrodo Red labelled Zymosan particles, without any adverse effects on neuron morphology. ioMicroglia start to fluoresce red when they have engulfed material which is initiated by a shift in pH.
Cells arrive ready to plate
Male donor-derived ioMicroglia 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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What scientists say about ioMicroglia
Matteo Zanella, PhD
Associate Research Leader | Charles River