Rapid and consistent generation of functional microglia from reprogrammed hiPSCs to study neurodegeneration and neuroinflammation
There is a need for a source of functional, consistent, scalable disease-relevant human microglia for neuroimmune research and the development of therapeutic or preventive strategies for neurodegeneration. This poster outlines the use of opti-ox precision cellular reprogramming for the rapid generation of homogenous and functionally consistent populations of ioMicroglia™ from human iPSCs in just 10 days. These microglia were shown to express key phenotypic markers, and RNA sequencing demonstrated that ioMicroglia have a transcriptomic signature similar to primary adult and foetal microglia. These microglia were also assessed for disease relevant functionality. ioMicroglia were shown to undergo phagocytosis of both E. coli particles and amyloid beta. Additionally when stimulated with LPS or IFNγ, ioMicroglia are shown to secrete pro-inflammatory cytokines. ioMicroglia can also be co-cultured with ioGlutamatergic Neurons™ to gain insights into complex cellular interactions.
In this poster, you will explore:
- How consistent, functional ioMicroglia can be generated from cryopreserved vials in just 10 days.
- Flow cytometry, and immunocytochemistry analysis of key microglia marker expression, including TMEM119, P2RY12, CD14, CD45 and CD11b.
- Phagocytosis assays showing that ioMicroglia undergo phagocytosis of E. coli and amyloid beta particles.
- Cytokine secretion assays showing that ioMicroglia secrete pro-inflammatory cytokines when exposed to LPS and IFNγ.
- Immunocytochemistry data showing ioMicroglia in co-culture with ioGlutamatergic Neurons.
- Single cell and bulk RNA-sequencing data showing that ioMicroglia are enriched for key phenotypic markers, and expression profiles closely cluster with primary adult and foetal microglia.
Raman et al