Derivation of Functional Human Astrocytes from Cerebral Organoids

Autor(es)

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

• Tipo de documento: Artigo Científico
• Fonte: SCIENTIFIC REPORTS; v. 7, MAR 27 2017.
• Citações Web of Science: 20

Resumo

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)

• Processo FAPESP: 14/21035-0 - Proteômica quantitativa de linhagens neurais e organóides cerebrais derivados de células tronco de pluripotência induzida de pacientes com esquizofrenia
• Beneficiário: Juliana Minardi Nascimento
• Modalidade de apoio: Bolsas no Brasil - Pós-Doutorado
• Processo FAPESP: 14/10068-4 - EMU concedido no processo 13/08711-3: espectrômetro de massas Waters SYNAPT G2-Si HDMs + nanoACQUITY UPLC
• Beneficiário: Daniel Martins-de-Souza
• Modalidade de apoio: Auxílio à Pesquisa - Programa Equipamentos Multiusuários
• Processo FAPESP: 13/08711-3 - Desenvolvimento de um teste preditivo para medicação bem sucedida e compreensão das bases moleculares da esquizofrenia através da proteômica
• Beneficiário: Daniel Martins-de-Souza
• Modalidade de apoio: Auxílio à Pesquisa - Jovens Pesquisadores
• Processo FAPESP: 16/07332-7 - O papel das vias de sinalização 14-3-3 e Efrina na comunicação celular entre neurônios-astrócitos e funcionamento da sinapse tripartite
• Beneficiário: Verônica Aparecida Monteiro Saia Cereda
• Modalidade de apoio: Bolsas no Brasil - Doutorado