Microglia are the primary immune cells in the central nervous system (CNS) and they play a crucial role in maintaining neuronal homeostasis and synaptic plasticity for normal brain development and neuronal function, and inflammatory responses. They have also been implicated in the pathogenesis of several neurological disorders such as Alzheimer’s disease and Parkinson’s disease. The differentiation of iPSCs to microglia provide a steady source of primary microglia-like cells without the sourcing issues associated with primary microglia. These cells are an excellent physiologically relevant research model to study immune response mechanisms in the brain, neuronal function, and for disease modeling.
Applied StemCell has developed an efficient cytokine-based method to differentiate high-quality microglia cells from human iPSCs (iMGLs), which recapitulate the phenotype and functional parameters of primary microglia and in vivo microglial cells. Our proprietary erythromyeloid induction protocol mimics the in vivo activation pathway for the development of microglia in the brain from hematopoietic stem cells.
The iMGLs were derived through directed differentiation from an integration-free, control human iPSC line (ASE-9211) reprogrammed from fibroblasts of an African-American male donor. These high-purity (≥90%) cells express microglial-specific markers TMEM119, and P2Ry12+ (also referred to as P2Y12+) as well as IBA1, CX3CR1+, and TREM2+ (Figure 1). The cells are provided as fully-differentiated, cryopreserved cells that you can readily recover and culture for your experiments. The iMGLs are functionally viable for 7 days after recovery.
To harness the full potential of our iMGLs, we also provide optimized, serum-free Microglia Culture Media (ASE-9601MM) that supports robust maintenance and functionality of the iMGLs in culture.
Contents of the Kit:
- iPSC-derived Microglia (iMGLs), ASE-9601-C
- Microglia Basal Culture Media, ASE-9601-MM-A
- Microglia Culture Media Supplement A, ASE-9601-MM-B
Characterization of the iMGL
Figure 1. Recovery of cryopreserved iPSC-derived microglia (iMGLs). Cryopreserved iMGLs, differentiated from Applied StemCell’s control iPSC line, ASE-9211 were recovered in microglia culture media. The cells were fixed the next day and stained with microglial-specific markers, P2RY12+, TMEM119, IBA1, CX3CR1, TREM2 and nuclear counterstain (DAPI). Images at 20x magnification.
Figure 2. iPSC-derived microglial cells were recovered and maintained in microglia growth media for 2 days before antibody staining for CD11b marker. Green: CD11b; Blue: DAPI.