cat no | io6015
ioMicroglia P2RY12 null/WT
Human iPSC-derived microglia model
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Cryopreserved human iPSC-derived cells powered by opti-ox, that are ready for functional experiments in 4 days
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Built to investigate the impact of P2RY12 knockout for neuroinflammation research
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Consistently perform key phagocytic and cytokine secretion functions, and are co-culture compatible
Human iPSC-derived microglia for neuroinflammation research
ioMicroglia Media Kit
Cell culture media kit for the culture of ioMicroglia up to 10 days post-thaw
Phagocytosis of ioMicroglia P2RY12null/wt is demonstrated comparably to the WT control and ioMicroglia P2RY12null/null
Phagocytosis was analysed at day 10 post-revival after incubation with 1 µg/0.33 cm2 pHrodo RED labelled E. coli particles for 24 hours +/- cytochalasin D control. The graph displays the proportion of cells phagocytosing E.coli over 24 hours. ioMicroglia P2RY12null/wt cells display a similar proportion of phagocytosis compared to the ioMicroglia P2RY12null/null and WT control. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. Three technical replicates were performed experiment.
Key cytokine secretion function displayed by ioMicroglia P2RY12 null/wt
Cytokine secretion was analysed at day 10 post-revival after stimulation with LPS 100 ng/ml and IFNɣ 20 ng/ml for 24 hours. This revealed that ioMicroglia P2RY12null/wt cells display a similar level of cytokine secretion compared to the ioMicroglia P2RY12null/null and WT control. Supernatants were harvested and analysed using MSD V-plex Proinflammatory Kit. Three technical replicates were performed per experiment.
ioMicroglia P2RY12 null/WT display clear ablation of ATP mediated chemotaxis
ioMicroglia Male (io1021), ioMicroglia P2RY12 null/WT, and ioMicroglia P2RY12 null/null (io6012) were plated onto Incucyte® Clearview 96-well Plate for Chemotaxis with 8 μm sized pores and 25 μM ATP in the basal compartment.
Cell movement through the pores onto the underside of the membrane was quantified with the Incucyte® S3 Live-Cell Analysis System using the Incucyte® Chemotaxis Analysis software (in collaboration with Sartorius).
A) The ratio of membrane coverage of the underside of the membrane to the top is plotted. N=6, two independent experiments, mean and SEM are shown.
B) Representative brightfield imaging of the underside of the transwell membranes at 62 h, blue shows the applied cell masking on each image.
As expected, clear ablation of ATP-mediated chemotaxis is seen both the ioMicroglia P2RY12 null/null and ioMicroglia P2RY12 null/WT cells.
ioMicroglia P2RY12 null/wt express IBA1 comparably to the genetically matched wild-type control
Immunofluorescent staining on day 10 post-revival demonstrates similar homogenous expression of the microglia marker IBA1 and ramified morphology in ioMicroglia P2RY12 null/wt cells compared to the genetically matched wild-type control, ioMicroglia Male. 100X magnification.
ioMicroglia P2RY12 null/wt show expected ramified morphology by day 10
ioMicroglia P2RY12null/wt cells mature rapidly and key ramified morphology can be identified by day 4 and continues through to day 10, similarly to the WT control. Day 1 to 10 post-thawing; 100x magnification.
P2RY12 null/wt heterozygous knockout confirmed by flow cytometry analysis
Flow cytometry analysis of ioMicroglia P2RY12 null/WT demonstrates heterozygous knockout of the P2RY12 gene translating to the protein level. Microglia purity demonstrated by >95% expression of CD45, CD11b and CD14 expression.
Phenotypic characterisation of a human iPSC-derived tri-culture using ioGlutamatergic Neurons, ioAstrocytes, and ioMicroglia
Using our fully optimised protocol, ioGlutamatergic Neurons (MAP2, red), ioMicroglia (IBA1, yellow) and ioAstrocytes (vimentin, cyan) were co-cultured to create a highly defined CNS model. High-resolution ICC analysis confirms the successful co-localisation and morphological health of three distinct cell types within a unified environment. By day 7, the protocol yields a highly consistent, integrated network suitable for complex cell modelling. DAPI (blue) highlights the total cell density and integrity of the culture. This protocol is compatible with derivative products of the three cell types, ensuring straightforward implementation across experimental workflows.
ioMicroglia are efficiently transfected with mRNA encoding GFP
ioMicroglia Male are efficiently transfected and show sustained long-term expression of mRNA encoding GFP. Cells were imaged throughout the experiment to assess transfection efficiency and evaluate potential cytotoxic effects of the transfection protocol. Day 4 images were captured prior to transfections on the same day.
Download the step-by-step protocol for lipid-based delivery of synthetic mRNA into ioMicroglia.
Vial limit exceeded
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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