cat no | io1035, io1036
ioMicroglia TREM2 R47H/R47H™
Human iPSC-derived Alzheimer's disease model
ioMicroglia TREM2 R47H/R47H are opti-ox™ precision reprogrammed microglia carrying a genetically engineered homozygous R47H mutation in the TREM2 gene encoding the triggering receptor expressed on myeloid cells 2 protein. The TREM2 R47H mutation has been linked with increased risk of late-onset Alzheimer's disease (AD).
These cells offer a functional, rapidly maturing, and disease relevant system to study the role of the TREM2 R47H mutation in late-onset AD, alongside a genetically matched wild-type control.
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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
Making True Comparisons
Pair the ioDisease Model Cells with the genetically matched wild-type ioMicroglia to directly investigate the effect of the mutant TREM2 protein on late-onset AD.
Quick
Rapidly maturing cells that are ready to use within 10 days post-revival, in mono- and co-cultures.
Functional
Disease model cells display key phagocytic and cytokine secretion functions.
Cells arrive ready to plate
ioMicroglia TREM2 R47H/R47H 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.
Highly characterised and defined
Disease model cells express key microglia markers comparably to the genetically matched wild-type control
Immunofluorescent staining on day 10 post-revival demonstrates similar homogenous expression of microglia markers P2RY12 and IBA1 and ramified morphology in both disease model clones compared to the genetically matched wild-type (WT) control. 100X magnification.
Disease model cells show expected ramified morphology by day 10
Both disease model clones 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.
Key phagocytic function
Disease model cells show a similar proportion of phagocytosis to the genetically matched wild-type control
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 particles over 24 hours and shows that both disease model clones display a similar proportion of phagocytosis to the WT control. Images were acquired every 30 mins on the Incucyte® looking at red fluorescence and phase contrast. Three technical replicates were performed per experiment.
Disease model cells show a similar degree of phagocytosis of E. coli particles to the genetically matched wild-type control
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 fluorescence intensity per cell displaying degree of phagocytosis per cell and shows that both disease model clones display a similar degree of phagocytosis per cell to the 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
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 both disease model clones secrete the predominantly pro-inflammatory cytokines, IL-6, IL-8, IL-1β, and TNF⍺ at a similar level to the WT control whereas IL-12p70 appears to be secreted at a lower level than the WT control. The anti-inflammatory IL-10 is secreted at a lower level by clone CL45 but at a higher level by clone CL17 than the WT control. Supernatants were harvested and analysed using MSD V-plex Proinflammatory Kit™. Three technical replicates were performed experiment.
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