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Human iPSC-Derived Microglia

Axol Human iPSC-Derived Microglia

iPSC-derived microglia

Axol’s Human iPSC-derived Microglia provide a reproducible, physiologically-relevant model in an easy-to-use, assay-ready format for investigating neuroglia involvement in neurodegeneration and neurodevelopment. We also offer a fully optimized serum-free Microglia Maintenance Medium to promote and support the maintenance of microglia.

Microglia are commonly described as the immune cells of the brain, with key roles in brain development, neurogenesis, synaptic plasticity and homeostatic maintenance.

Axol’s method for generating iPSC-derived microglia mimics the in vivo pathway of development for brain resident macrophages and produces microglia that are representative of primary human microglia in vitro. Our iPSC-derived Microglia deliver the advantage that they provide an almost infinite source of microglia from a single donor with a normal karyotype.

The homogenous and reproducible population of our iPSC-derived Microglia exhibit physiologically relevant functionality as they are highly phagocytic and produce anti-inflammatory factors in response to pathogens. Our iPSC-derived Microglia also express the microglia-specific marker TMEM119 along with myeloid markers TREM2 and IBA-1. These phenotypes make iPSC-derived Microglia suitable models for investigating neuroinflammation in Alzheimer’s disease, multiple sclerosis and Parkinson’s disease.

Axol’s Human iPSC-derived Microglia are coming soon! Register your interest below to be first to find out about our release date!


iPSC-derived microglia

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Microglia culture media and reagents

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Morphological characterization

Brightfield images of Axol’s Human iPSC-Derived Microglia show typical morphology with long ramified processes characteristic of human microglia.

Morphological characterization of Axol Human iPSC-Derived Microglia

Brightfield images of Axol’s Human iPSC-Derived Microglia. Human iPSC-derived monocytes were seeded using Microglia Maintenance Medium into 96-well plates at a density of 100,000 cells/cm2. During maturation the spherical monocytes adhere strongly to the culture surface and as they differentiate to microglia their morphology becomes ramified. Axol’s iPSC-Derived Microglia show surveillance behaviour which is representative of their phenotype in vivo.


Immunocytochemistry

Human iPSC-derived Microglia show protein expression of myeloid and microglia-specific markers IBA-1, TREM2 and TMEM119.

Immunocytochemistry of Human iPSC-derived Microglia after 14 days in culture

Immunocytochemistry of Human iPSC-derived Microglia, after 14 days in culture, revealed the expression of TMEM119, a specific microglia transmembrane protein which enables the explicit identification of brain resident microglia. Central nervous system myeloid markers, TREM2 and IBA-1 were also expressed.


Phagocytosis assay

Phagocytosis was used to measure functional activation and inhibition of Human iPSC-derived Microglia. Phagocytic activity was assessed using pHrodo green Zymosan bioparticles and Human iPSC-derived Microglia were found to be highly phagocytic.

Phagocytosis analysis of human iPSC-derived Microglia co-cultured with human iPSC-derived Cortical Neurons

The iPSC-derived microglia were measured using a fluorescence plate reader, (excitation 480nm/ emission 530nm; 20nm filter). Values were normalized to background fluorescence using excitation and emission wavelengths of 300nm and 530nm, respectively.

Axol’s Human iPSC-derived Microglia are highly phagocytic

Human iPSC-derived Microglia (ax0666) were cultured for 14 days before stimulating with 100ng/ml LPS for 1 hour or treated with 10 μM Cytochalasin D (CytoD) for 30 minutes prior to adding pHrodo green Zymosan particles at 0.15 mg/mL. Phagocytosis was allowed to progress for 1 hour and 30 minutes before fixation with PFA.

The presence of Zymosan particles induced an inflammatory response in the Human iPSC-derived Microglia which, was able to be significantly increased by lipopolysaccharide (LPS). In turn, CytoD significantly inhibited the uptake of Zymosan particles into microglia.


Cytokine response

Axol’s Human iPSC-derived Microglia secrete cytokines in response to inflammatory stimuli.

Cytokine response of Axol Human iPSC-Derived Microglia

Secretion of the pro-inflammatory cytokine, Tumour Necrosis Factor – alpha (TNF-α), was measured from the cellular supernatants of Axol’s Human iPSC-derived Microglia and Human iPSC-derived Macrophages, using an AlphaLISA. Both microglia and macrophages, from the same donor, showed secretion of TNF-α exhibiting physiological function.

Data kindly provided by Sygnature Discovery.


Co-culture of microglia and cortical neurons

Axol’s Human iPSC-derived Microglia are suitable for co-culture with Human iPSC-derived Cortical Neurons.

Immunocytochemistry of Human iPSC-derived Microglia co-cultured with Human iPSC-derived cortical neurons

A. Immunocytochemistry of Human iPSC-derived Microglia co-cultured with Human iPSC-derived cortical neurons reveals microglia specific maker expression TMEM119 and neuronal marker β-3 Tubulin.

Phagocytosis analysis of human iPSC-derived Microglia co-cultured with human iPSC-derived Cortical Neurons
Cytokine response of human iPSC-derived Microglia co-cultured with human iPSC-derived Cortical Neurons