Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation
Summary: 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) ag...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-03-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S266616672500084X |
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| author | Rosa González-Sastre Raquel Coronel Patricia Mateos-Martínez Laura Maeso Elena Llorente-Beneyto Alfonso Luque Berta Anta Victoria López-Alonso Isabel Liste |
| author_facet | Rosa González-Sastre Raquel Coronel Patricia Mateos-Martínez Laura Maeso Elena Llorente-Beneyto Alfonso Luque Berta Anta Victoria López-Alonso Isabel Liste |
| author_sort | Rosa González-Sastre |
| collection | DOAJ |
| description | Summary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-bbcca994812b4aaa8d32a924d49f853d |
| institution | DOAJ |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-03-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-bbcca994812b4aaa8d32a924d49f853d2025-08-20T02:47:40ZengElsevierSTAR Protocols2666-16672025-03-016110367810.1016/j.xpro.2025.103678Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregationRosa González-Sastre0Raquel Coronel1Patricia Mateos-Martínez2Laura Maeso3Elena Llorente-Beneyto4Alfonso Luque5Berta Anta6Victoria López-Alonso7Isabel Liste8Unidad de Regeneración Neural, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Unidad de Biología Computacional, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Doctoranda en la Escuela Internacional de Doctorado de la Universidad Nacional de Educación a Distancia (UNED), Programa en Ciencias Biomédicas y Salud PúblicaUnidad de Regeneración Neural, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Corresponding authorUnidad de Regeneración Neural, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Unidad de Biología Computacional, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Doctoranda en la Escuela Internacional de Doctorado de la Universidad Nacional de Educación a Distancia (UNED), Programa en Ciencias Biomédicas y Salud PúblicaUnidad de Biología Computacional, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, SpainUnidad de Regeneración Neural, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Doctoranda en la Escuela Internacional de Doctorado de la Universidad Nacional de Educación a Distancia (UNED), Programa en Ciencias Biomédicas y Salud PúblicaUnidad de Endotelio Funcional, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, SpainUnidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, SpainUnidad de Biología Computacional, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Corresponding authorUnidad de Regeneración Neural, Unidad Funcional de Investigación de Enfermedades Crónicas (UFIEC), Instituto de Salud Carlos III (ISCIII), Majadahonda, Madrid 28220, Spain; Corresponding authorSummary: 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.1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S266616672500084XNeuroscienceStem CellsOrganoids |
| spellingShingle | Rosa González-Sastre Raquel Coronel Patricia Mateos-Martínez Laura Maeso Elena Llorente-Beneyto Alfonso Luque Berta Anta Victoria López-Alonso Isabel Liste Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation STAR Protocols Neuroscience Stem Cells Organoids |
| title | Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation |
| title_full | Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation |
| title_fullStr | Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation |
| title_full_unstemmed | Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation |
| title_short | Protocol for generating human cerebral organoids from two-dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation |
| title_sort | protocol for generating human cerebral organoids from two dimensional cultures of pluripotent stem cells bypassing embryoid body aggregation |
| topic | Neuroscience Stem Cells Organoids |
| url | http://www.sciencedirect.com/science/article/pii/S266616672500084X |
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