title: Derivation of Functional Human Astrocytes from Cerebral Organoids.
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.
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ., Brasil.
authors.forename: Rômulo Sperduto
authors.initials: RS
authors.lastname: Dezonne
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ., Brasil.|Instituto D'Or de Pesquisa e Ensino (IDOR), Rio de Janeiro, RJ, Brasil.
authors.forename: Rafaela Costa
authors.initials: RC
authors.lastname: Sartore
authors.affiliation: Instituto D'Or de Pesquisa e Ensino (IDOR), Rio de Janeiro, RJ, Brasil.|Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brasil.
authors.forename: Juliana Minardi
authors.initials: JM
authors.lastname: Nascimento
authors.affiliation: Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brasil.
authors.forename: Verônica M
authors.initials: VM
authors.lastname: Saia-Cereda
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ., Brasil.|Universidade Federal do Rio de Janeiro,Campus Xerém, RJ, Brasil.
authors.forename: Luciana Ferreira
authors.initials: LF
authors.lastname: Romão
authors.affiliation: Hospital Universitário Clementino Fraga Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
authors.forename: Soniza Vieira
authors.initials: SV
authors.lastname: Alves-Leon
authors.affiliation: Hospital Universitário Clementino Fraga Filho, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ, Brasil.
authors.forename: Jorge Marcondes
authors.initials: JM
authors.lastname: de Souza
authors.affiliation: Instituto de Biologia, Universidade Estadual de Campinas, Campinas, SP, Brasil.
authors.forename: Daniel
authors.initials: D
authors.lastname: Martins-de-Souza
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ., Brasil.|Instituto D'Or de Pesquisa e Ensino (IDOR), Rio de Janeiro, RJ, Brasil.
authors.forename: Stevens Kastrup
authors.initials: SK
authors.lastname: Rehen
authors.affiliation: Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, RJ., Brasil.
authors.forename: Flávia Carvalho Alcantara
authors.initials: FC
authors.lastname: Gomes
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000378:metabolism; D020013:Calcium Signaling; D002478:Cells, Cultured; D002540:Cerebral Cortex / Q000166:cytology*; D003598:Cytoskeletal Proteins / Q000235:genetics / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006801:Humans; D051379:Mice; D000071446:Neuronal Outgrowth; D009940:Organoids / Q000166:cytology*; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D020543:Proteome / Q000235:genetics / Q000378:metabolism
chemical_list: D003598:Cytoskeletal Proteins; D020543:Proteome; D018698:Glutamic Acid