cat no | io1029
ioMicroglia Female
Female human iPSC donor-derived microglia
ioMicroglia (io1029) are female donor-derived human microglial cells from iPSC, deterministically programmed using opti-ox technology.
Within 10 days post-revival, ioMicroglia are ready for experimentation, expressing (>90%) key microglia markers, including 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 female 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 deterministic programmed ioMicroglia from a female donor rapidly form a homogenous microglia population.
Time-lapse video capturing the rapid and homogeneous microglia phenotype acquisition upon thawing of cryopreserved female donor-derived ioMicroglia. 10 day time course.
Highly characterised and defined
Flow cytometry analysis of female donor-derived ioMicroglia shows key phenotypic marker expression
Flow cytometry analysis of day 10 female donor-derived ioMicroglia shows key microglia marker expression of CD11b, CD45, CD14, and P2RY12 with a purity of above 97% for all these markers.
View the cell detachment protocol used to generate this data.
Female donor-derived ioMicroglia show key microglia marker expression
Immunofluorescent staining of day 10 female donor-derived ioMicroglia shows homogenous expression of P2RY12, IBA1 and TREM2, and a typical ramified morphology. DAPI counterstain (blue). Image taken at 10x magnification.
Female donor-derived ioMicroglia show ramified morphology by day 10
Rapid morphological changes in the female donor-derived 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.
Whole transcriptome analysis demonstrates high lot-to-lot consistency of female donor-derived ioMicroglia
Bulk RNA sequencing analysis was performed on three independent lots of female donor-derived ioMicroglia at three different time points throughout the reprogramming protocol. Principal component analysis represents the variance in gene expression between the lots and shows the high consistency across each lot at each given time point. Populations of female donor-derived ioMicroglia with equivalent expression profiles can be generated consistently from every vial, allowing confidence in experimental reproducibility.
Whole transcriptome analysis demonstrates that female donor-derived ioMicroglia are highly similar to primary adult, foetal and other iPSC-derived microglia
Principal component analysis of bulk RNA sequencing data from female 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
Female donor-derived ioMicroglia display a different level of phagocytosis than male donor-derived cells
Day 10 female donor-derived ioMicroglia (io1029) from three independent lots and male donor-derived ioMicroglia (io1021) from one lot were incubated with pHrodo RED labelled Zymosan particles for 24 hours +/- cytochalasin D control. The graph displays that the proportion of cells phagocytosing Zymosan particles over 24 hours is consistent across three independent lots and that female donor-derived ioMicroglia cells display a higher proportion of phagocytosis than male donor-derived cells. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. Three technical replicates were performed per lot.
Female donor-derived ioMicroglia display a consistent degree of phagocytosis across lots
Day 10 female donor-derived ioMicroglia (io1029) from three independent lots and male donor-derived ioMicroglia (io1021) from one lot were incubated with pHrodo™ RED labelled Zymosan particles for 24 hours +/- cytochalasin D control. The graph displays that the degree of cells phagocytosing Zymosan particles over 24 hours is consistent across three independent lots. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. Three technical replicates were performed per lot.
Female donor-derived ioMicroglia phagocytose Zymosan particles
Representative video showing female donor-derived ioMicroglia (io1029) phagocytosing pHrodo Red labelled Zymosan particles. When female donor-derived ioMicroglia engulf these particles this causes the particles to fluoresce red, within the cells, due to the drop in pH in the phagolysosome. Live imaging was performed in 2-minute intervals over a time period of 2 hours using the 3D Cell Explorer 96focus Nanolive Imaging system.
Female donor-derived ioMicroglia display a different pro-inflammatory cytokine response to male donor-derived cells
Co-culture compatible
ioGlutamatergic Neurons (io1001) were cultured to day 10 post-thaw. Female donor-derived ioMicroglia (io1029) 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 6 days. Representative video showing that female donor-derived ioMicroglia form a stable co-culture with ioGlutamatergic Neurons. Live imaging was performed in 6.5-minute intervals over a time period of 3 hours and 31 minutes using the 3D Cell Explorer 96focus Nanolive Imaging system.
Key marker expression in female donor-derived ioMicroglia and ioGlutamatergic Neuron co-cultures
Immunofluorescent analysis at day 8 of the co-cultures shows expression of the microglia marker, IBA1 (green) and the pan-neuronal marker, MAP2 (red), as expected. Representative images taken at 10x magnification.
Female donor-derived ioMicroglia retain phagocytic function in co-culture with ioGlutamatergic Neurons
Representative video showing female-derived donor ioMicroglia (io1029) in co-culture with ioGlutamatergic Neurons (io1001) selectively phagocytosing pHrodo Red labelled Zymosan particles after 10 days in co-culture, without any observed adverse effects on neuron morphology. When female donor-derived ioMicroglia engulf these particles this causes the particles to fluoresce red, within the cells, due to the drop in pH in the phagolysosome. Live imaging was performed in 8-minute intervals over a time period of 1 hour and 36 minutes using the 3D Cell Explorer 96focus Nanolive Imaging system.
Cells arrive ready to plate
Female 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