Protocol for generating human cortical organoids enriched in outer radial glia by guided differentiation
The generation of human pluripotent stem cell (hPSC)-derived brain organoids is continuously refined, enhancing their reproducibility and complexity. Here, we present a guided differentiation protocol for generating cortical forebrain organoids and cortico-pericyte (CP)assembloids composed of a robust outer radial glia (oRG) population and an expanded outer subventricular zone (oSVZ). We describe the steps to generate hPSC-derived cortical organoids (COs), cortical pericytes, and CP assembloids. Moreover, we outline the procedures to characterize the organoids by immunostaining and to perform single-cell dissociation. For complete details on the use and execution of this protocol, please refer to Walsh et al.<sup>1</sup>.
Authors
- Ryan Walsh (ORCID: https://orcid.org/0000-0003-2338-3115)
- Raffaele Luongo
- Arianna Baggiolini (ORCID: https://orcid.org/0000-0002-2476-561X)
- Lorenz Studer (ORCID: https://orcid.org/0000-0003-0741-7987)
- Antonietta Verrillo (ORCID: https://orcid.org/0009-0002-1131-8439)
Institutions
- Memorial Sloan Kettering Cancer Center (US)
- Cornell University (US)
- Università della Svizzera italiana (CH)
Publication Details
- Journal
- STAR Protocols
- Published
- 2024-08-29
- DOI
- https://doi.org/10.1016/j.xpro.2024.103280
- Citations
- 1
- Primary Topic
- Pluripotent Stem Cells Research
- Type
- article
- Field-Weighted Citation Impact
- 0.23
Funders
- U.S. Department of Defense
- Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung
- National Institutes of Health