PT  - JOURNAL ARTICLE
AU  - Vukasin M. Jovanovic
AU  - Claire Malley
AU  - Carlos A. Tristan
AU  - Seungmi Ryu
AU  - Pei-Hsuan Chu
AU  - Elena Barnaeva
AU  - Pinar Ormanoglu
AU  - Jennifer Colon Mercado
AU  - Sam Michael
AU  - Michael E. Ward
AU  - Anton Simeonov
AU  - Ilyas Singeç
TI  - Directed Differentiation of Human Pluripotent Stem Cells into Radial Glia and Astrocytes Bypasses Neurogenesis
AID  - 10.1101/2021.08.23.457423
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.08.23.457423
4099  - http://biorxiv.org/content/early/2021/08/23/2021.08.23.457423.short
4100  - http://biorxiv.org/content/early/2021/08/23/2021.08.23.457423.full
AB  - Derivation of astrocytes from human pluripotent stem cells (hPSCs) is inefficient and cumbersome, impeding their use in biomedical research. Here, we developed a highly efficient chemically defined astrocyte differentiation strategy that overcomes current limitations. This approach largely bypasses neurogenesis, which otherwise precedes astrogliogenesis during brain development and in vitro experiments. hPSCs were first differentiated into radial glial cells (RGCs) exhibiting in vivo-like radial glia signatures. Activation of NOTCH and JAK/STAT pathways in bona fide RGCs resulted in direct astrogliogenesis confirmed by expression of various glial markers (NFIA, NFIB, SOX9, CD44, S100B, GFAP). Transcriptomic and genome-wide epigenetic analyses confirmed RGC-to-astrocyte differentiation and absence of neurogenesis. The morphological and functional identity of hPSC-derived astrocytes was confirmed by using an array of methods (e.g. electron microscopy, calcium imaging, co-culture with neurons, grafting into mouse brains). Lastly, the scalable protocol was adapted to a robotic platform and used to model Alexander disease. In conclusion, our findings uncover remarkable plasticity in neural lineage progression that can be exploited to manufacture large numbers of human hPSC-derived astrocytes for drug development and regenerative medicine.Competing Interest StatementV.M.J., A.S., and I.S. are co-inventors on a US Department of Health and Human Services patent application covering the RGC and astrocyte differentiation method and its use.