ioMicroglia TREM2 R47H Het hero image compressed FINAL2

cat no | io1038

ioMicroglia TREM2 R47H/WT

Human iPSC-derived Alzheimer's disease model

ioMicroglia TREM2 R47H/WT are opti-ox precision reprogrammed microglia carrying a genetically engineered heterozygous 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.

Place your order

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.

per vial

Benchtop benefits

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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.

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Quick

Rapidly maturing cells that are ready to use within 10 days post-revival, in mono- and co-cultures.

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Functional

Disease model cells display key phagocytic and cytokine secretion functions.

Technical data

Cells arrive ready to plate

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ioMicroglia TREM2 R47H/WT 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
ioMicroglia TREM2 R47H Het P2RY12 panel FINAL
ioMicroglia TREM2 R47H Het IBA1 panel FINAL

Immunofluorescent staining on day 10 post-revival demonstrates similar homogenous expression of microglia markers P2RY12 and IBA1 and ramified morphology in the disease model clone compared to the genetically matched wild-type (WT) control. 100X magnification.

Disease model cells show expected ramified morphology by day 10
ioMicroglia TREM2 R47H Het Morphology panel final compressed

The disease model clone matures 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 reduced proportion of phagocytosis of E.coli particles compared to the genetically matched wild-type control

ioMicroglia TREM2 R47H Het phago prop ecoli graph

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 the disease model clone displays a reduced proportion of phagocytosis than 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 reduced degree of phagocytosis of E. coli particles compared to the genetically matched wild-type control

ioMicroglia TREM2 R47H Het phago intensity ecoli graph

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 the disease model clone displays a reduced degree of phagocytosis per cell than 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. 

Key cytokine secretion function

Disease model cells display reduced secretion of  pro-inflammatory cytokines upon activation compared to the genetically matched wild-type control

ioMicroglia TREM2 R47H Het cytokine graphs FINAL

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 the disease model clone secretes the predominantly pro-inflammatory cytokines, IL-6, IL-8, IL-10, IL-12p70, IL-1β, and TNF⍺ at a lower level than the WT control. Supernatants were harvested and analysed using MSD V-plex Proinflammatory Kit. Three technical replicates were performed per experiment. 

Product information

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

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

Genetic modification

Heterozygous R47H mutation in the TREM2 gene

Applications

Alzheimer's disease modelling
Drug discovery and development
Neuroinflammation modelling
Phagocytosis assays
Cytokine response assays
Co-culture studies

Available clones

io1038S: ioMicroglia TREM2 R47H/WT (CL86)

Product resources

Improving physiological relevance in neurological disease drug development Case study
Improving physiological relevance in neurological disease drug development

Elise Malavasi, PhD
Principal Scientist
Concept Life Sciences

Download
Quantifying C5a-mediated chemotaxis in precision reprogrammed hiPSC-derived ioMicroglia Application note
Quantifying C5a-mediated chemotaxis in precision reprogrammed hiPSC-derived ioMicroglia

bit.bio | Medicines Discovery Catapult

2024

Download
CRISPR knockout screening for drug target identification and validation using CRISPR-Ready ioMicroglia Poster
CRISPR knockout screening for drug target identification and validation using CRISPR-Ready ioMicroglia
Byrne et al. 
bit.bio
2024
Download
ioMicroglia Brochure
ioMicroglia
bit.bio
Download

Alzheimer’s Disease Pathogenesis - Emerging Role of Microglia

In this GEN webinar, hear from our distinguished expert, Dr Matthias Pawlowski, and learn about the emerging role of microglia in the pathogenesis of Alzheimer’s disease and their potential as a therapeutic target to treat this disease effectively.

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