Project description:Protocol to differentiate iPSC control lines into astrocytes. We performed the RNA sequencing of cells at different time point: embryoid bodies ; neurospheres at passage 1 ; neurospheres at passage 2 and differentiated astrocytes (iPast) for the 201B7 iPSC and iPast stage for WD39.
Project description:The goals of this study are to generate inflammation-sensitive astrocytes from human induced pluripotent stem cells. We examine the transcriptomic inflammatory signature of generated astrocytes following Il1Beta exposure. Primary human cerebellar astrocytes, human induced pluripotent stem cells (hiPSC)-derived neural precursor cells (NPCs) and hiPSC-derived astrocytes were treated with Il1beta and compared to vehicle treated controls. Results: hiPSC-derived can be differentiated to astrocytes that are inflammation sensitive.
Project description:The ability to generate astrocytes from human pluripotent stem cells (hPSCs) offers a promising cellular model to study the development and physiology of human astrocytes. However, the extant methods for generating functional astrocytes required long culture periods and remained much ambiguity whether their paradigm follows the innate developmental program. In this report, we provided an efficient and rapid method for generating physiologically functional astrocytes from hPSCs. Overexpressing the nuclear factor IB (NFIB) in hPSC-derived neural precursor cells (NPCs) induced a highly enriched astrocyte population in 2 weeks. RNA sequencing and functional analyses demonstrated progressive transcriptomic and physiological changes in the cells, resembling in vivo astrocyte development. Further analyses substantiated previous results and established the MAPK pathway necessary for astrocyte differentiation. Hence, this differentiation paradigm provides a prospective in vitro model for human astrogliogenesis studies and pathophysiology of neurological diseases concerning astrocytes.
Project description:In this study, we present evidence for the first time of senescent astrocytes derived from human pluripotent stem cells (hPSCs) induced by an extended culture.
Project description:Recent studies highlighted the importance of astrocytes in neuroinflammatory diseases, interacting closely with other CNS cells but also with the immune system. However, due to the difficulty in obtaining human astrocytes, their role in these pathologies is still poorly characterized. Here, we develop a new serum-free protocol to differentiate human iPSCs into astrocytes. Gene expression and functional assays show that our protocol consistently yields a highly enriched population of resting mature astrocytes across the thirteen hiPSC lines differentiated. Using this new model, we first highlight the importance of serum-free media for astrocyte culture to generate resting astrocytes. Second, we assess the astrocytic response to IL-1β, TNFα and IL-6, all cytokines important in neuroinflammation, such as multiple sclerosis. Our study reveals very specific profiles of reactive astrocytes depending on the triggering stimulus. This new model provides ideal conditions for in-depth and unbiased characterization of astrocyte reactivity in neuroinflammatory conditions.