Deriving Functional Astrocytes from Mouse Embryonic Stem Cells with a Fast and Efficient Protocol

A growing number of studies highlight the structural and functional diversity of astrocytes throughout the central nervous system. These cells are now seen as heterogeneous as neurons and are implicated in a number of neurological and psychiatric diseases. Efficient generation of diverse subtypes of astrocytes can be a useful tool in investigating synaptogenesis and patterns of activity in developing neural networks. In this study, we developed a protocol for the fast and efficient differentiation of astrocytes from mouse embryonic stem cells, as evidenced by the upregulation of genes related to astrocytic development (Gfap, Aldh1l1). Generated astrocytes exhibit phenotypic diversity, which is demonstrated by the variant expression of markers such as GFAP, ALDH1L1, AQP4 and S100β, amongst subgroups within the same cell population. In addition, astrocytes exhibited differential calcium transients upon stimulation with ATP. Our protocol will facilitate investigations, regarding the involvement of astrocytes in the structural and functional connectivity of neural networks.