ioMicroglia TREM2 R47H Hom Hero image compressed FINAL

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.

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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/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
ioMicroglia TREM2 R46H Hom P2RY12 ICC panel FINAL compressed
ioMicroglia TREM2 R46H Hom IBA1 ICC panel FINAL compressed 2

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
ioMicroglia TREM2 R47H Hom Morphology panel FINAL compressed

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

ioMicroglia TREM2 R47H Hom phago prop ecoli_FINAL

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

ioMicroglia TREM2 R47H Hom phago intensity ecoli_FINAL

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

Disease model cells secrete pro-inflammatory cytokines upon activation
ioMicroglia TREM2 R47H Hom 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 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. 

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

Homozygous 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

io1035S: ioMicroglia TREM2 R47H/R47H (CL17)
io1036S: ioMicroglia TREM2 R47H/R47H (CL45)

Product resources

Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer’s disease Poster
Generation and characterisation of a panel of human iPSC-derived neurons and microglia carrying early and late onset relevant mutations for Alzheimer’s disease
Smith et al. 
bit.bio
2024
Downlaod
ioMicroglia™ and related disease models User manual
ioMicroglia™ and related disease models

V6

bit.bio

2023

Download
Mastering Cell Identity In A Dish: The Power Of Cellular Reprogramming Webinar
Mastering Cell Identity In A Dish: The Power Of Cellular Reprogramming

Prof Roger Pedersen | Adjunct Professor and Senior Research Scientist at Stanford University 

Dr Thomas Moreau | Director of Cell Biology Research | bit.bio

Watch now
Rethinking Developmental Biology With Cellular Reprogramming Webinar
Rethinking Developmental Biology With Cellular Reprogramming

Mark Kotter | CEO and founder | bit.bio

Marius Wernig | Professor Departments of Pathology and Chemical and Systems Biology |  Stanford University

Watch now

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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