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Application Note

Characterization of human iPSC-derived Macrophages reveals the hallmarks of macrophage morphology and function

The use of macrophages from peripheral blood or immortalized cell lines may be reducing the reproducibility and reliability of your disease research and its successful translation into new treatments.


In vitro characterization of human induced pluripotent stem cell-derived macrophages

Human induced pluripotent stem cell-derived macrophages (hiPSCmacrophages) (Axol Bioscience) offer a physiologically relevant model for modelling infection and immuno-oncology screening in vitro.

hiPSC-macrophages are a type off immune cell (leukocyte or white blood cell) differentiated from hiPSC-derived monocytes. Macrophages play an essential role in phagocytosis, immunity and have an anti-inflammatory role in a variety of tissue types throughout the body. Macrophages are, thus, able to clear up cell debris, microbes and foreign substances through engulfing or internalising and destroying them by a process referred to as acidification. Moreover, when recruited to the site of injury or infection, macrophages have the ability to differentiate into tissue specific macrophages such as microglia, which are part of the nervous system.

Here we present data on the differentiation and characterization of hiPSCmacrophages. We determined seeding parameters and optimal differentiation conditions of hiPSC-derived monocytes to hiPSC-derived macrophages. The hiPSC-derived macrophages were, subsequently, assessed for expression of specific macrophage cell surface markers , using a cytokine array. Phagocytosis was then assessed using zymosan particles.

Here we have demonstrated that hiPSC-macrophage offer a suitable, reliable and reproducible alternative to human blood derived macrophages for, not only, infection and immuno-oncology studies, but also for co-culture studies for tissue regeneration and repair studies.


Development of high content approaches for investigating microglia function in neurodegenerative disorders

The role of Central Nervous System (CNS) resident macrophages, termed microglia, in neurodegeneration have come to prominence in recent years, being important in brain homeostasis and the clearance of pathogenic material. Microglial function has been implicated in a wide variety of CNS disease including Amyloid Lateral Sclerosis (ALS), Alzheimer’s, Huntington’s and Parkinson’s Disease (AD, HD & PD respectively) and Multiple Sclerosis (MS).

Accessing primary human cells, whether monocyte, macrophage or microglia, in sufficient quantities for drug discovery is extremely challenging and so a number of immortalized human monocyte cell lines and primary material from murine and rodent sources are available to act as a surrogate of the phagocytic ability of human microglial cells. We have established robust high content assays in these cell types to assess phagocytic function with the use of various fluorescent substrates, these have been successfully multiplexed with quantification of target proteins and cell viability assessments. These assays are HTS compatible and are capable of being performed in 1,536 well plates, allowing conservation of precious primary material and expensive fluorescent reagents.

More recently, iPSC derived monocyte and macrophage cells have become commercially available for use within these assays. As these can be obtained from patients with underlying CNS diseases, they may provide information on therapeutics which can modify some of the phenotypic markers of CNS disease.

Here we describe a selection of the assays we have established to study phagocytic activity as a surrogate for microglial function.


iPSC-Derived Macrophage Protocol

Cell culture protocol for maintaining Axol Human iPSC-Derived Macrophages in a 96 well plate format.