Abstract
Single-cell mRNA sequencing can dissect heterogeneous cell populations as it can identify cell types and cellular states based on their unique transcriptional signatures. We use fluorescence-activated cell sorting (FACS) to isolate individual cultured neurons derived from human-induced pluripotent stem cells (hiPSCs) followed by polyA-based Smart-Seq2 RNA sequencing to analyze the single-cell transcriptional profiles. We provide protocols and guidelines on dissociation, cell selection, and library preparation that can be readily adapted to other cell types or tissue samples.
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Acknowledgments
We thank Mattias Karlen for help with Fig. 1. Flow cytometry was performed in the Biomedicum Flowcytometry Core Facility with support of the Karolinska Institutet. We thank Juan Basile and Belinda Pannagel for assisting with flow cytometry. We are grateful for discussions with other members of the Hedlund laboratory. We are also thankful for discussions with and support from members of Rickard Sandberg’s lab. The work in the Hedlund laboratory is supported by grants from the Swedish Research Council (grant number 2020-01049), The Radala Foundation for ALS Research (Switzerland), Ulla-Carin Lindquists Foundation for ALS Research (Ulla-Carin Lindquists stiftelse för ALS forskning), Åhlén-stiftelsen (grant number 213051), Olav Thon Stiftelsen (Norway), The Swedish Brain Foundation (Hjärnfonden) (grant number FO2021-0145) and Parkinsonfonden (grant number 1328/21). C.S. was supported by an Early Postdoc.Mobility fellowship from the Swiss National Science Foundation (P2BEP3_172233).
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Schweingruber, C., Nijssen, J., Benitez, J.A., Hedlund, E. (2023). Single-Cell mRNA-Seq of In Vitro-Derived Human Neurons Using Smart-Seq2. In: Song, Q., Tao, Z. (eds) Transcription Factor Regulatory Networks. Methods in Molecular Biology, vol 2594. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2815-7_11
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DOI: https://doi.org/10.1007/978-1-0716-2815-7_11
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