cat no | io6012
ioMicroglia P2RY12 null/null
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
Phagocytosis capability of ioMicroglia P2RY12 null/null is comparable to the WT control and ioMicroglia P2RY12 null/WT
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 P2RY12 null/null cells display a similar proportion of phagocytosis compared to the ioMicroglia P2RY12 null/WT 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/null
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 P2RY12 null/null cells display a similar level of cytokine secretion compared to the ioMicroglia P2RY12 null/WT and WT control. Supernatants were harvested and analysed using MSD V-plex Proinflammatory Kit. Three technical replicates were performed per experiment.
ioMicroglia P2RY12 null/null 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/null cells compared to the genetically matched wild-type control, ioMicroglia Male. 100X magnification.
ioMicroglia P2RY12 null/null show expected ramified morphology by day 10
ioMicroglia P2RY12 null/null 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/null homozygous knockout confirmed by flow cytometry analysis
Flow cytometry analysis of ioMicroglia P2RY12 null/null demonstrates homozygous knockout of the P2RY12 gene translating to the protein level. Microglia purity demonstrated by >95% expression of CD45, CD11b and CD14 expression.
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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