Derivation of Functional Human Astrocytes from Cerebral Organoids

Author(s)

Dezonne, Romulo Sperduto ; Sartore, Rafaela Costa ; Nascimento, Juliana Minardi ; Saia-Cereda, Veronica M. ; Romao, Luciana Ferreira ; Alves-Leon, Soniza Vieira ; de Souza, Jorge Marcondes ; Martins-de-Souza, Daniel ; Rehen, Stevens Kastrup ; Alcantara Gomes, Flvia Carvalho

• Document type: Journal article
• Source: SCIENTIFIC REPORTS; v. 7, MAR 27 2017.
• Web of Science Citations: 20

Abstract

Astrocytes play a critical role in the development and homeostasis of the central nervous system (CNS). Astrocyte dysfunction results in several neurological and degenerative diseases. However, a major challenge to our understanding of astrocyte physiology and pathology is the restriction of studies to animal models, human post-mortem brain tissues, or samples obtained from invasive surgical procedures. Here, we report a protocol to generate human functional astrocytes from cerebral organoids derived from human pluripotent stem cells. The cellular isolation of cerebral organoids yielded cells that were morphologically and functionally like astrocytes. Immunolabelling and proteomic assays revealed that human organoid-derived astrocytes express the main astrocytic molecular markers, including glutamate transporters, specific enzymes and cytoskeletal proteins. We found that organoid-derived astrocytes strongly supported neuronal survival and neurite outgrowth and responded to ATP through transient calcium wave elevations, which are hallmarks of astrocyte physiology. Additionally, these astrocytes presented similar functional pathways to those isolated from adult human cortex by surgical procedures. This is the first study to provide proteomic and functional analyses of astrocytes isolated from human cerebral organoids. The isolation of these astrocytes holds great potential for the investigation of developmental and evolutionary features of the human brain and provides a useful approach to drug screening and neurodegenerative disease modelling. (AU)

• FAPESP's process: 14/21035-0 - Quantitative proteomics in neural cell lines and organoids derived from induced pluripotent stem cells from schizophrenia patients
• Grantee: Juliana Minardi Nascimento
• Support Opportunities: Scholarships in Brazil - Post-Doctoral
• FAPESP's process: 14/10068-4 - Multi-User Equipment approved in grant 13/08711-3: mass spectrometer waters SYNAPT G2-Si HDMS + nanoACQUITY UPLC
• Grantee: Daniel Martins-de-Souza
• Support Opportunities: Multi-user Equipment Program
• FAPESP's process: 13/08711-3 - Developing a predictive test for a successful medication response and understanding the molecular bases of schizophrenia through proteomics
• Grantee: Daniel Martins-de-Souza
• Support Opportunities: Research Grants - Young Investigators Grants
• FAPESP's process: 16/07332-7 - The role of 14-3-3 and Ephrin signaling pathways on cellular communication between neurons-astrocytes and the functioning of the synapse tripartite
• Grantee: Verônica Aparecida Monteiro Saia Cereda
• Support Opportunities: Scholarships in Brazil - Doctorate