cat no | ioA021 Early Access
ioMicroglia are human induced pluripotent stem cell (iPSC)-derived microglia, precision reprogrammed using opti-ox™ technology. Within 10 days post-revival, ioMicroglia are ready for experimentation, expressing (>90%) key phenotypic markers, including TMEM119, P2RY12, IBA1, CX3CR1, CD11b, CD45, and CD14. ioMicroglia display key functional traits including phagocytosis and proinflammatory cytokine secretion, and can be co-cultured with ioGlutamatergic Neurons™ with retention of phagocytic function.
ioMicroglia provide a rapid, consistent, functional, and easy-to-use hiPSC-based model for neurodegenerative disease research and drug development.
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Quick
Rapidly maturing human microglia that are ready to use within 10 days post-revival.
Functional
ioMicroglia display key phagocytic and cytokine secretion functions.
Co-culture compatible
Suitable for co-culture studies with neurons.
Starting material
Human iPSC line
Seeding compatibility
6, 12, 24 & 96 well plates
Shipping info
Dry ice
Donor
Caucasian adult male (skin fibroblast)
Vial size
Small: >1.5 x 10⁶ viable cells
Quality control
Sterility, protein expression (ICC), 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
ioMicroglia generated by transcription factor-driven reprogramming of iPSCs using opti-ox technology
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.
ICC analysis of ioMicroglia shows key phenotypic marker expression
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; 400µm.
Principal component analysis (PCA) of bulk RNA sequencing data from ioMicroglia, integrated with sequencing data from Abud et al. (1) and Chen et al. (2), show that ioMicroglia cluster closely to primary foetal and adult microglia data sets derived from both these publications. 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
(2) Chen SW, et al., Stem Cell Reports, 2021; 16(5): 1363-1380
Day 10 ioMicroglia 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. Graph displays the proportion of cells phagocytosing E. coli particles over 24 hours. Three technical replicates were performed per batch.
Day 10 ioMicroglia 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 two independent batches. Three technical replicates were performed per batch.
ioMicroglia form co-cultures with ioGlutamatergic Neurons
Video showing cryopreserved ioMicroglia and ioGlutamatergic Neurons that were thawed and cultured independently for 10 days before being co-cultured for 8 days. Imaging was then performed in 30-minute intervals. ioMicroglia form a stable co-culture with ioGlutamatergic Neurons.
Key marker expression in ioMicroglia and ioGlutamatergic Neuron co-cultures
ioMicroglia retain phagocytic function in co-culture with ioGlutamatergic Neurons
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.
Mark Kotter | CEO and founder | bit.bio
Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology | Stanford University
2023
V1
bit.bio
2022
University of Münster & bit.bio
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.