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细胞培养基及添加物

STEMdiff™ Microglia Maturation Kit

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

概要
技术资料
数据及文献

概要

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.

数据及文献

Data

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.

RNA-seq profiles of hPSC-derived microglia and other immune cell types are plotted after principal components analysis.

Figure 2. Microglia Generated with STEMdiff™ Microglia Culture System Are Transcriptionally Similar to Those from Published Differentiation and Maturation Protocols

RNA-seq datasets were extracted from 4 different publications (protocols A-D) that generated hPSC-derived microglia and compared their transcriptional profiles to those of other immune cell types (N = 500 genes). Principal component analysis (PCA) was performed on these data along with RNA-seq data from microglia generated with STEMdiff™ Microglia Culture System. The hPSC-derived microglia from STEMdiff™ Microglia Culture System plot most closely to those from protocols A and B.

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

Figure 3. 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 4. 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.