Single-Cell Functional Analysis of Human Astrocytes: How IsoPlexis’ Newest Protocol is Expanding Neuroscience Research Capabilities

Neuroinflammation has been implicated in the pathogenesis and progression of many neurological disorders. To uncover drivers of inflammation, is it critical to interrogate the function of individual immune cells to analyze how cells contribute to the overall immune response. IsoPlexis has developed and validated various protocols for analysis of cells of the central nervous system, including monocyte-derived dendritic cells (MoDCs), monocyte-derived microglia-like cells (MDMi cells), and mouse astrocytes, to empower researchers to better understand the heterogenous landscape of immune cells in the brain. To expand upon central nervous system cell profiling capabilities, IsoPlexis has developed a new protocol to functionally phenotype human induced pluripotent stem cell derived (iPSC-derived) astrocytes.

Though initially believed to be solely support cells, astrocytes have been discovered to be crucial for facilitating several functions that restore and maintain key mechanisms within the brain, including impacting brain plasticity, supporting neuronal metabolic processes, and repairing the brain following injury or disease. As astrocytes are constantly changing to accomplish these crucial functions, it is important to characterize how these cells facilitate immune responses to better understand how neurological disorders and injuries progress and how new therapies can be developed to treat them.

IsoPlexis’ Human Astrocyte Protocol Overview

IsoPlexis’ IsoCode Single-Cell Adaptive Immune: Human Induced Pluripotent Stem Cell Derived (iPSC-derived) Astrocyte Protocol provides researchers with a validated workflow to analyze human astrocyte cells using the IsoPlexis platform. First, cryopreserved cells are thawed and cultured. Next, the iPSC-derived astrocytes are stimulated with TNF-α, IL-1α, and C1q before staining. Following staining with Cell Stain 405, the cells are lifted from the plates and stained once more with FITC CD49f, then loaded onto the IsoCode chips for automated single-cell analysis by the IsoLight or IsoSpark instrument.

Like all of IsoPlexis’ application protocols, the ELISA-based workflow is fully automated with our instruments, the IsoLight and IsoSpark. IsoPlexis’ integrated software analysis platform, IsoSpeak, empowers researchers to make sense of high-dimensional proteomics data easily and rapidly with the click of a button – no coding or specialized personnel required.

Explore the Brain’s Immune Landscape with Advanced Single-Cell Applications

The new Human iPSC-derived Astrocyte Protocol adds to the wide range of immune cell types that can be functionally phenotyped using IsoPlexis’ platform. IsoPlexis provides unique insights into all stages of the research pipeline from discovery to clinical development, helping researchers uncover key biomarkers to better understand disease and develop new therapies. To see all of IsoPlexis’ cell profiling capabilities, check out our Applications Protocols page, which showcases the workflows and cell types that have been internally tested and validated on the IsoPlexis platform.

Learn more about how leading researchers are using single-cell functional proteomics to combat the world’s toughest diseases in our newest roundtable eBook, Single-Cell Advances in Functional Immune Monitoring for Developing Immune Therapies and Uncovering Disease Pathogenesis. Download below!

 

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