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iCell Microglia AD TREM2, 01279
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Kits
- Cryopreserved iCell Microglia TREM2 (HZ), 01279
- 50ml iCell Glial Base Medium
- 0.5ml iCell Microglia Supplement A (100x)
- 0.5ml iCell Microglia Supplement B (100x)
- 1ml iCell Neural Supplement C (50X)
- iCell Microglia Quick Guide
- Additional reagents required; see Quick Guide
genetic status
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Catalog #:
Microglia differentiated from human iPS cells, frozen
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$1,995.00
Cells Only
- Cryopreserved iCell Microglia TREM2 (HZ), 01279
- iCell Microglia Quick Guide
- Additional reagents required; see Quick Guide
genetic status
Quantity (Cells Per Vials)
Catalog #
Catalog #:
Microglia differentiated from human iPS cells, frozen
From
$1,695.00
Product Overview
Microglia are immune cells resident in the central nervous system (CNS) responsible for fundamental physiological and pathological processes. Microglia support neuronal homeostasis and facilitate neuronal network formation by synaptic pruning. Surveillant microglia can activate, adapt and respond accordingly to specific stimuli.
Microglia dysfunction has been implicated as a key player in neurodegeneration afflictions, including Alzheimer’s Disease (AD). The limited availability and inconsistency of primary human microglia has constrained research and therapeutic progress.
Rare variants of TREM2 (triggering receptor expressed on myeloid cells 2), an immune receptor expressed on microglia known to trigger phagocytosis of bacteria and the release of reactive oxygen species, are associated with an increased risk of developing AD. Most TREM2 mutations found in AD risk variants are heterozygous mutations that impact the binding of TREM2 ligands or the shedding of the extracellular domains. Loss‐of‐function mutations of TREM2 and adaptor protein TYROBP have also been linked to the development of Nasu–Hakola disease, an inflammatory degenerative disease of the brain and bone, leading to premature dementia and death.
Engineered TREM2 Models
To enable investigation of the functional consequences of TREM2 variants, a frameshift in TREM2 exon 2 in either one allele (heterozygous; C60MfsTer45; iCell Microglia TREM2 HZ) or both alleles (homozygous; P59AfsTer16 and P59AfsTer46; iCell Microglia TREM2 HO) near amino acid 60 was engineered in iPSCs from an otherwise apparently healthy, normal donor background, 01279.
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Technical Docs
PROTOCOLS
Application Protocols
Immunofluorescent Labeling of iCell Microglia
Application Protocols
Labeling Amyloid Beta with pHrodo Red
Application Protocols
Measuring Microglia Phagocytosis: Kinetic Imaging on the IncuCyte
Application Protocols
Tri-Culture of iCell Microglia with iCell Neuronal Cell Types and iCell Astrocytes
USER DOCS
Quick Guide
iCell Microglia Quick Guide
Datasheet
iCell Microglia
Biosafety Document
iCell Microglia Kit, TREM2 (HT), 01279
Biosafety Document
iCell Microglia Kit, TREM2 (HO), 01279
Biosafety Document
iCell Microglia Biosafety Document
Safety Data Sheet
iCell Neural Supplement C (50X) Safety Data Sheet - Asia
Safety Data Sheet
iCell Neural Supplement C (50X) Safety Data Sheet
Safety Data Sheet
iCell Glial Basal Medium Safety Data Sheet - Asia
Safety Data Sheet
iCell Glial Basal Medium Safety Data Sheet
Safety Data Sheet
iCell Neural Supplement C (50X) Safety Data Sheet
Safety Data Sheet
iCell Neural Supplement C (50X) Safety Data Sheet - Asia
Safety Data Sheet
iCell Cell-Based Products Safety Data Sheet - Asia
Safety Data Sheet
iCell Cell-Based Products Safety Data Sheet
Safety Data Sheet
iCell Glial Base Medium Safety Data Sheet
Safety Data Sheet
iCell Glial Base Medium Safety Data Sheet - Asia
Safety Data Sheet
iCell Neural Supplement C Safety Data Sheet
Safety Data Sheet
iCell Neural Supplement C Safety Data Sheet - Asia
Safety Data Sheet
iCell Microglia Supplement A Safety Data Sheet
Safety Data Sheet
iCell Microglia Supplement A Safety Data Sheet - Asia
Safety Data Sheet
iCell Microglia Supplement B Safety Data Sheet
Safety Data Sheet
iCell Microglia Supplement B Safety Data Sheet - Asia
Performance Data
Comparative Marker Expression
Comparative Marker Expression iCell Microglia were labeled for the presence of cell surface (CD45, CD11b, CD11c, TREM2 and, CD33) and intracellular (P2RY12, TMEM119, CX3CR1, IBA1) antigens by flow cytometry. The specific staining is compared against matched isotype controls. A comparison of cell surface and intracellular antigens reveals similar marker expression across iCell Microglia 01279 and the two TREM2 engineered lines (Fig. 1).
Figure 1: Comparison of Marker Expression in iCell Microglia and the Engineered Variants
Figure 2: Phagocytosis of iCell Microglia and TREM2 Variants
Phagocytosis
Phagocytosis In Figure 2, a comparison is shown between iCell Microglia and the engineered TREM2 lines. Phagocytosis of S.aureus BioParticles® was measured over time using an IncuCyte® S3 Live-Cell Analysis System. The iCell Microglia TREM2 HZ and iCell Microglia HO exhibit phagocytic activity, but at a reduced activity relative to the control line. The reduced phagocytosis in TREM2 variants were correlated to their survival in vitro (data not shown).
TREM2 Expression Levels
TREM2 Expression Levels Soluble TREM2 levels were quantified from the iCell Microglia control and TREM2 variants using a SimpleStep ELISA (Fig. 3A). The TREM2 antibody recognizes the extracellular portion of the TREM2 protein. The iCell Microglia control and TREM2 variants were thawed and plated at the same density in maintenance media in a 96-well plate. The spent media was collected at 3 days and 7 days post thaw to determine the levels of soluble TREM2. The amount of TREM2 in total protein lysates was measured by Western blot. Cell lysates of iCell Microglia control, TREM2 and another engineered line, iCell Microglia SNCA A53T, were electrophoretically separated and detected with anti-TREM2 antibody using the ProteinSimple Wes System along with a molecular weight markers (Fig. 3B).
Figure 3: Measurement of TREM2 Protein by ELISA and Western Blot
Figure 4: Cytokine Comparison of iCell Microglia and TREM2 Variants
Cytokine Response
Cytokine Response The cytokine response of stimulated iCell Microglia and TREM2 variants were compared by multiplex assay (Fig. 4). iCell Microglia were plated in maintenance media and allowed to recover for 72 h prior to stimulation with LPS, interferon gamma or both for 24 h. Supernatants were collected and assayed using the multiplex Luminex system.
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Product Highlights
Schematic Representation of TREM2 Protein
Figure 1. Schematic Representation of TREM2 Protein
A frameshift mutation engineered in exon 2 of the TREM2 gene (indicated by the arrow) interferes with the signal peptide domain.
Figure 2. Engineered Variants of 01279 and the Predicted Amino Acid Sequence for the TREM2 Exon 2 Domain
Engineered Variants of 01279 and the Predicted Amino Acid Sequence for the TREM2 Exon 2 Domain
Advantages
- Human relevance: iCell Microglia exhibit functional characteristics similar to human microglia, including phagocytosis and cytokine-mediated inflammatory responses.
- Reproducible research: Rigorous quality control and large-scale manufacturing capabilities ensure consistent performance from every lot of iCell Microglia (>80% TREM2+, IBA1+, CX3CR1+).
- Rapid results: iCell Microglia are fully differentiated cells (not precursors) allowing for more experiments in less time.
- Disease lines available: TREM2 and MeCP2 engineered mutants and isogenic control.