STEMdiff™ Microglia Maturation Kit

Maturation kit for the generation of microglia from human ES and iPS cell-derived microglia precursors

STEMdiff™ Microglia Maturation Kit

Maturation kit for the generation of microglia from human ES and iPS cell-derived microglia precursors

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Maturation kit for the generation of microglia from human ES and iPS cell-derived microglia precursors
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Product Advantages


  • Greater than 90% of source HPCs are differentiated into microglia.

  • Upstream compatibility with HPCs generated using the STEMdiff™ Hematopoietic Kit and co-culture with STEMdiff™ neural organoids.

  • Allows differentiation to microglia with few contaminating macrophages or monocytes.

  • Microglia generated with the kit demonstrate phagocytosis and activation.

  • Required materials are provided in a simple, easy-to-use format.

What's Included

  • STEMdiff™ Microglia Maturation Kit (Catalog #100-0020)
    • STEMdiff™ Microglia Basal Medium, 90 mL
    • STEMdiff™ Microglia Supplement 1, 10 mL
    • STEMdiff™ Microglia Supplement 2, 400 uL
    • STEMdiff™ Microglia Supplement 3, 400 uL
Products for Your Protocol
To see all required products for your protocol, please consult the Protocols and Documentation.

Overview

The STEMdiff™ Microglia Culture System comprises STEMdiff™ Microglia Differentiation Kit and STEMdiff™ Microglia Maturation Kit. Together, these kits are used to differentiate and mature microglia derived from human pluripotent stem cells (hPSCs) using STEMdiff™ Hematopoietic Kit (Catalog #05310).

Based on the protocol from the laboratory of Mathew Blurton-Jones (Abud et al., 2017), the resulting cells are a highly pure population of microglia (> 80% CD45/CD11b-positive, > 50% TREM2-positive microglia; < 20% morphologically distinct monocytes or macrophages).

Cells derived using these products are versatile tools for modeling neuroinflammation, studying human neurological development and disease, co-culture applications, and toxicity testing.
Subtype
Specialized Media
Cell Type
Hematopoietic Cells, PSC-Derived, Microglia, Neural Cells, PSC-Derived
Species
Human
Application
Cell Culture, Differentiation
Brand
STEMdiff
Area of Interest
Disease Modeling, Drug Discovery and Toxicity Testing, Immunology, Neuroscience
Formulation Category
Serum-Free

Data Figures

Experimental protocol schematic for generating microglial precursors from hPSC-derived hematopoietic progenitor cells.

Figure 1. Schematic for the STEMdiff™ Microglia Culture System Protocol

Functional microglia can be generated in 4-10 days from microglial precursors after 24 days in STEMdiff™ Microglia Differentiation Medium. For the differentiation of microglial precursors from hPSC-derived hematopoietic progenitor cells, see the PIS.

Bulk RNA-seq heatmap showing that STEMdiff™ Microglia express disease-relevant genes similar to those from popular protocols.

Figure 2. Microglia Generated with STEMdiff™ Microglia Culture System Express Disease-Relevant Genes Similar to Those from Published Differentiation and Maturation Protocols

Bulk RNA-seq datasets were extracted from 8 different publications that generated hPSC- (iMGL) and primary- (MGL) derived microglia and their transcriptional profiles compared to data from microglia generated with STEMdiff™ Microglia Culture System. The heat map displays absolute expression levels for select genes associated with Alzheimer’s disease, Parkinson’s disease, and viral encephalitis. Significant differences in gene expression between microglia generated with STEMdiff™ Microglia Culture System and any of the other 3 groups were identified by differential gene expression analysis. *= p<0.05 (DEseq2, adjusted). hPSC = human pluripotent stem cell.

Graphs demonstrating that STEMdiff™ Microglia release cytokines in response to LPS stimulation, compared to control.

Figure 3. Microglia Generated with STEMdiff™ Microglia Culture System Release Cytokines in Response to Inflammatory Signals

Microglia were generated using the STEMdiff™ Microglia Culture System and stimulated with 100 ng/mL LPS for 24 hours. The release of pro-inflammatory (TNFα, IL-6, IFN-γ, IL-1β, GM-CSF, IL-12p70, IL-2, IL-8) and anti-inflammatory (IL-10) cytokines were measured by MSD. The microglia release cytokines in response to LPS treatment, as expected. *, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ****, p ≤ 0.0001. LPS = lipopolysaccharide; MSD = Meso Scale Discovery.

Time lapse of functional microglia phagocytosing bioindicator particles in culture. Over time, cells turn red and aggregate.

Figure 4. STEMdiff™ Microglia Culture System Generates Functional Microglia Capable of Phagocytosis at Day 34

Microglia taking up pH-sensitive bioindicator particles at a concentration of 250 μg/mL (small dots) were measured over a 12-hour time period with live cell imaging. As the particles are phagocytosed, the cells turn red. Over time, the number of small dots decreased, and the red cells increased in number and aggregated. Scale bar = 100μm.

Figure 5. PSC-Derived Microglia Incorporate into Brain Organoids After 10 Days and Display an Activated Morphology upon Injury.

(A) Representative microglia and brain organoid co-cultures after 10 days, stained with IBA1 for microglia (green) and MAP2 for neurons (magenta). The microglia integrate among the neurons and display an unactivated morphology with extended processes (arrow). (B) The microglia display an activated amoeboid morphology upon injury as shown by IBA1 staining.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
100-0020
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
100-0020
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
100-0020
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
100-0020
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
100-0020
Lot #
All
Language
English

Applications

This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.

Resources and Publications

Educational Materials (19)