cat no | io1021
ioMicroglia are human microglial cells precision reprogrammed from iPSC, 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.
ioMicroglia recapitulate key human microglia functions with lot-to-lot consistency, including mediating an inflammatory response, disposal of unwanted materials, and carrying out immune surveillance. In addition, ioMicroglia also display chemotaxis and can be co-cultured with ioGlutamatergic Neurons™ to gain insights into complex intercellular interactions.
ioMicroglia provide a functional, consistent, rapid, and easy-to-use hiPSC-based model for neurodegenerative disease research and drug development.
per vial
A maximum number of 20 vials applies. If you would like to order more than 20 vials, please contact us at orders@bit.bio.
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Functional
ioMicroglia display key phagocytic and cytokine secretion functions with lot-to-lot consistency.
Quick
Rapidly maturing 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.
opti-ox precision reprogrammed ioMicroglia rapidly form a homogenous microglia population.
Time-lapse video capturing the rapid and homogeneous microglia phenotype acquisition upon thawing of cryopreserved ioMicroglia. 10 day time course.
Flow cytometry analysis of ioMicroglia shows key phenotypic marker expression
Flow cytometry analysis of day 10 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.
Immunofluorescent staining of day 10 ioMicroglia shows homogenous expression of P2RY12, IBA1 and TREM2, and a typical ramified morphology. DAPI counterstain (blue). Image taken at 10x magnification.
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; 10x magnification; scale bar; 400 µm.
Principal component analysis of bulk RNA sequencing data from ioMicroglia, integrated with sequencing data from Abud et al. (1) shows that ioMicroglia 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
Day 10 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.
Day 10 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.
ioGlutamatergic Neurons were cultured to day 10 post-thaw. ioMicroglia 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 ioMicroglia form a stable co-culture with ioGlutamatergic Neurons.
Key marker expression in 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.
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 four-phase process: Phase 0: 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.
Starting material
Human iPSC line
Seeding compatibility
6, 12, 24, 96 & 384 well plates
Shipping info
Dry ice
Donor
Caucasian adult male (skin fibroblast)
Vial size
Small: >1.5 x 10⁶ viable cells
Large: >5 x 10⁶ viable cells
Quality control
Sterility, protein expression (ICC), functional phagocytosis and cytokine secretion assays
Differentiation method
opti-ox cellular reprogramming
Recommended seeding density
37,000 to 39,500 cells/cm²
User storage
LN2 or -150°C
Format
Cryopreserved cells
Product use
ioCells are for research use only
Applications
Disease modeling
Drug development
Neuroinflammation research
Co-culture studies
Transcriptome analysis
Prof Roger Pedersen | Adjunct Professor and Senior Research Scientist at Stanford University
Dr Thomas Moreau | Director of Cell Biology Research | bit.bio
V4
bit.bio
2023
Mark Kotter | CEO and founder | bit.bio
Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology | Stanford University
Madeleine Garrett | Field Application Specialist | bit.bio
An interview with a leading researcher and microglia expert Dr Anthony Vernon at King's College Institute of Psychiatry, Psychology & Neuroscience, to demystify the complex roles of microglia in our brand new blog.