title: MicroRNA-based conversion of human fibroblasts into striatal medium spiny neurons.
abstract: The ability to generate human neurons of specific subtypes of clinical importance offers experimental platforms that may be instrumental for disease modeling. We recently published a study demonstrating the use of neuronal microRNAs (miRNAs) and transcription factors to directly convert human fibroblasts to a highly enriched population of striatal medium spiny neurons (MSNs), a neuronal subpopulation that has a crucial role in motor control and harbors selective susceptibility to cell death in Huntington's disease. Here we describe a stepwise protocol for the generation of MSNs by direct neuronal conversion of human fibroblasts in 30 d. We provide descriptions of cellular behaviors during reprogramming and crucial steps involved in gene delivery, cell adhesion and culturing conditions that promote cell survival. Our protocol offers a unique approach to combine microRNAs and transcription factors to guide the neuronal conversion of human fibroblasts toward a specific neuronal subtype.
authors.affiliation: Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Michelle
authors.initials: M
authors.lastname: Richner
authors.affiliation: Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri, USA.|Program in Neuroscience, Division of Biology and Biomedical Sciences, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Matheus B
authors.initials: MB
authors.lastname: Victor
authors.affiliation: Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Yangjian
authors.initials: Y
authors.lastname: Liu
authors.affiliation: Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri, USA.|Program in Developmental, Regenerative and Stem Cell Biology, Division of Biology and Biomedical Sciences, Washington University, St. Louis, Missouri, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Abernathy
authors.affiliation: Department of Developmental Biology, Washington University School of Medicine, St. Louis, Missouri, USA.
authors.forename: Andrew S
authors.initials: AS
authors.lastname: Yoo
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D005347:Fibroblasts / Q000166:cytology*; D006801:Humans; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000166:cytology*
chemical_list: D035683:MicroRNAs