Differentiation of functional astrocytes from human-induced pluripotent stem cells in chemically defined media

Summary This protocol describes how to obtain human astrocytes from human-induced pluripotent stem cells (hiPSCs) in chemically defined media, without the use of fetal bovine serum (FBS). FBS eases the differentiation of astrocytes but also deeply alters their phenotype, as compared with their in vivo counterparts. Our protocol generates hiPSC-derived astrocytes displaying a phenotype and functions similar to human primary astrocytes, including adequate response to inflammation, neurotransmitter uptake, and trophic support to neurons. For complete details on the use and execution of this protocol, please refer to Perriot et al. (2018).
Graphical abstract
Highlights • Differentiation of astrocytes from hiPSCs in chemically defined media • Generation of highly characterized resting human astrocytes • Full functionality of hiPSC-derived astrocytes as compared with primary cells • Suitable model to study human astrocyte responses to inflammation
This protocol describes how to obtain human astrocytes from human-induced pluripotent stem cells (hiPSCs) in chemically defined media, without the use of fetal bovine serum (FBS). FBS eases the differentiation of astrocytes but also deeply alters their phenotype, as compared with their in vivo counterparts. Our protocol generates hiPSC-derived astrocytes displaying a phenotype and functions similar to human primary astrocytes, including adequate response to inflammation, neurotransmitter uptake, and trophic support to neurons.