title: Direct conversion of fibroblasts into functional astrocytes by defined transcription factors.
abstract: Direct cell reprogramming enables direct conversion of fibroblasts into functional neurons and oligodendrocytes using a minimal set of cell-lineage-specific transcription factors. This approach is rapid and simple, generating the cell types of interest in one step. However, it remains unknown whether this technology can be applied to convert fibroblasts into astrocytes, the third neural lineage. Astrocytes play crucial roles in neuronal homeostasis, and their dysfunctions contribute to the origin and progression of multiple human diseases. Herein, we carried out a screening using several transcription factors involved in defining the astroglial cell fate and identified NFIA, NFIB, and SOX9 to be sufficient to convert with high efficiency embryonic and postnatal mouse fibroblasts into astrocytes (iAstrocytes). We proved both by gene-expression profiling and functional tests that iAstrocytes are comparable to native brain astrocytes. This protocol can be then employed to generate functional iAstrocytes for a wide range of experimental applications.
authors.affiliation: Stem Cell and Neurogenesis Unit, Division of Neuroscience, San Raffaele Scientific Institute, Milan 20132, Italy. Electronic address: massimiliano.caiazzo@epfl.ch.
authors.forename: Massimiliano
authors.initials: M
authors.lastname: Caiazzo
authors.affiliation: Stem Cell and Neurogenesis Unit, Division of Neuroscience, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Serena
authors.initials: S
authors.lastname: Giannelli
authors.affiliation: Section of Physiology, Department of Experimental Medicine, University of Genoa and National Institute of Neuroscience, 16132 Genoa, Italy.
authors.forename: Pierluigi
authors.initials: P
authors.lastname: Valente
authors.affiliation: Department of Neuroscience and Brain Technologies, Italian Institute of Technology, 16132 Genoa, Italy.
authors.forename: Gabriele
authors.initials: G
authors.lastname: Lignani
authors.affiliation: Telethon Institute of Genetics and Medicine, Naples 80131, Italy.
authors.forename: Annamaria
authors.initials: A
authors.lastname: Carissimo
authors.affiliation: Stem Cell and Neurogenesis Unit, Division of Neuroscience, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Sessa
authors.affiliation: Stem Cell and Neurogenesis Unit, Division of Neuroscience, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Gaia
authors.initials: G
authors.lastname: Colasante
authors.affiliation: Telethon Institute of Genetics and Medicine, Naples 80131, Italy; Dulbecco Telethon Institute.
authors.forename: Rosa
authors.initials: R
authors.lastname: Bartolomeo
authors.affiliation: Stem Cell and Neurogenesis Unit, Division of Neuroscience, San Raffaele Scientific Institute, Milan 20132, Italy.
authors.forename: Luca
authors.initials: L
authors.lastname: Massimino
authors.affiliation: Department of Pharmacy and Biotechnology, University of Bologna, Bologna 40126, Italy.
authors.forename: Stefano
authors.initials: S
authors.lastname: Ferroni
authors.affiliation: Telethon Institute of Genetics and Medicine, Naples 80131, Italy; Dulbecco Telethon Institute; Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, TX 77030, USA; Jan and Dan Duncan Neurological Research Institute, Texas Children's Hospital, Houston, TX 77030, USA; Medical Genetics, Department of Medical and Translational Science Unit, Federico II University, Via Pansini 5, 80131 Naples, Italy.
authors.forename: Carmine
authors.initials: C
authors.lastname: Settembre
authors.affiliation: Section of Physiology, Department of Experimental Medicine, University of Genoa and National Institute of Neuroscience, 16132 Genoa, Italy; Department of Neuroscience and Brain Technologies, Italian Institute of Technology, 16132 Genoa, Italy.
authors.forename: Fabio
authors.initials: F
authors.lastname: Benfenati
authors.affiliation: Stem Cell and Neurogenesis Unit, Division of Neuroscience, San Raffaele Scientific Institute, Milan 20132, Italy. Electronic address: broccoli.vania@hsr.it.
authors.forename: Vania
authors.initials: V
authors.lastname: Broccoli
mesh_terms: D000818:Animals; D001253:Astrocytes / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015415:Biomarkers; D054338:Cell Transdifferentiation / Q000187:drug effects / Q000235:genetics*; D002478:Cells, Cultured; D065150:Cellular Reprogramming / Q000235:genetics; D016000:Cluster Analysis; D016207:Cytokines / Q000378:metabolism / Q000494:pharmacology; D005347:Fibroblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D015870:Gene Expression; D020869:Gene Expression Profiling; D006801:Humans; D008564:Membrane Potentials / Q000187:drug effects / Q000235:genetics; D051379:Mice; D010641:Phenotype; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism
chemical_list: D015415:Biomarkers; D016207:Cytokines; D014157:Transcription Factors