Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation
Human cerebral organoids (hCOs) provide an excellent model for the study of human brain development and disease. Here, we present a protocol to obtain hCOs directly from two-dimensional (2D) pluripotent stem cell (PSC) cultures, avoiding cell dissociation and posterior embryoid body (EB) aggregation. We describe steps for subjecting 2D cultures to a neural fate and subsequently developing hCOs. We then detail the evaluation of different cellular types. For complete details on the use and execution of this protocol, please refer to González-Sastre et al.<sup>1</sup>.
Authors
- B. Anta (ORCID: https://orcid.org/0000-0001-5972-5485)
- Victoria López-Alonso
- Patricia Mateos-Martínez (ORCID: https://orcid.org/0000-0001-9293-9100)
- Raquel Coronel (ORCID: https://orcid.org/0000-0002-1156-2151)
- Rosa González-Sastre
- Laura Maeso
- Isabel Liste
- Alfonso Luque
- Elena Llorente-Beneyto
Institutions
- Unidad Funcional de Investigación de Enfermedades Crónicas (ES)
Publication Details
- Journal
- STAR Protocols
- Published
- 2025-03-01
- DOI
- https://doi.org/10.1016/j.xpro.2025.103678
- Citations
- 1
- Primary Topic
- Pluripotent Stem Cells Research
- Type
- article
- Field-Weighted Citation Impact
- 0.81
Funders
- Ministerio de Ciencia, Innovación y Universidades
- Ministerio de Ciencia e Innovación
- Instituto de Salud Carlos III
- European Regional Development Fund
- Agencia Estatal de Investigación