Lineage recording in human cerebral organoids.

Autor: He Z; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Maynard A; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Jain A; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Gerber T; Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany., Petri R; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Lin HC; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland.; Institute of Molecular and Clinical Ophthalmology, Basel, Switzerland., Santel M; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Ly K; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Dupré JS; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Sidow L; Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany., Sanchis Calleja F; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Jansen SMJ; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland., Riesenberg S; Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany., Camp JG; Institute of Molecular and Clinical Ophthalmology, Basel, Switzerland. jarrettgrayson.camp@unibas.ch.; Department of Ophthalmology, University of Basel, Basel, Switzerland. jarrettgrayson.camp@unibas.ch., Treutlein B; Department of Biosystems Science and Engineering, ETH Zürich, Basel, Switzerland. barbara.treutlein@bsse.ethz.ch.; Max Planck Institute for Evolutionary Anthropology, Leipzig, Germany. barbara.treutlein@bsse.ethz.ch.
Jazyk: angličtina
Zdroj: Nature methods [Nat Methods] 2022 Jan; Vol. 19 (1), pp. 90-99. Date of Electronic Publication: 2021 Dec 30.
DOI: 10.1038/s41592-021-01344-8
Abstrakt: Induced pluripotent stem cell (iPSC)-derived organoids provide models to study human organ development. Single-cell transcriptomics enable highly resolved descriptions of cell states within these systems; however, approaches are needed to directly measure lineage relationships. Here we establish iTracer, a lineage recorder that combines reporter barcodes with inducible CRISPR-Cas9 scarring and is compatible with single-cell and spatial transcriptomics. We apply iTracer to explore clonality and lineage dynamics during cerebral organoid development and identify a time window of fate restriction as well as variation in neurogenic dynamics between progenitor neuron families. We also establish long-term four-dimensional light-sheet microscopy for spatial lineage recording in cerebral organoids and confirm regional clonality in the developing neuroepithelium. We incorporate gene perturbation (iTracer-perturb) and assess the effect of mosaic TSC2 mutations on cerebral organoid development. Our data shed light on how lineages and fates are established during cerebral organoid formation. More broadly, our techniques can be adapted in any iPSC-derived culture system to dissect lineage alterations during normal or perturbed development.
(© 2021. The Author(s).)
Databáze: MEDLINE
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