title: Efficient and cost-effective generation of mature neurons from human induced pluripotent stem cells.
abstract: For years, our ability to study pathological changes in neurological diseases has been hampered by the lack of relevant models until the recent groundbreaking work from Yamanaka's group showing that it is feasible to generate induced pluripotent stem cells (iPSCs) from human somatic cells and to redirect the fate of these iPSCs into differentiated cells. In particular, much interest has focused on the ability to differentiate human iPSCs into neuronal progenitors and functional neurons for relevance to a large number of pathologies including mental retardation and behavioral or degenerative syndromes. Current differentiation protocols are time-consuming and generate limited amounts of cells, hindering use on a large scale. We describe a feeder-free method relying on the use of a chemically defined medium that overcomes the need for embryoid body formation and neuronal rosette isolation for neuronal precursors and terminally differentiated neuron production. Four days after induction, expression of markers of the neurectoderm lineage is detectable. Between 4 and 7 days, neuronal precursors can be expanded, frozen, and thawed without loss of proliferation and differentiation capacities or further differentiated. Terminal differentiation into the different subtypes of mature neurons found in the human brain were observed. At 6-35 days after induction, cells express typical voltage-gated and ionotrophic receptors for GABA, glycine, and acetylcholine. This specific and efficient single-step strategy in a chemically defined medium allows the production of mature neurons in 20-40 days with multiple applications, especially for modeling human pathologies.
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Cherif
authors.initials: C
authors.lastname: Badja
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Galyna
authors.initials: G
authors.lastname: Maleeva
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Claire
authors.initials: C
authors.lastname: El-Yazidi
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Emilie
authors.initials: E
authors.lastname: Barruet
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Manon
authors.initials: M
authors.lastname: Lasserre
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Philippe
authors.initials: P
authors.lastname: Tropel
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Bernard
authors.initials: B
authors.lastname: Binetruy
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA.
authors.forename: Piotr
authors.initials: P
authors.lastname: Bregestovski
authors.affiliation: Medical Genetics and Functional Genomics and Brain Dynamics Institute, Aix-Marseille University, INSERM, Marseille, France; University of California San Francisco, San Francisco, California, USA frederique.magdinier@univ-amu.fr.
authors.forename: Frédérique
authors.initials: F
authors.lastname: Magdinier
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D058953:Neural Stem Cells / Q000166:cytology / Q000378:metabolism*; D009474:Neurons / Q000166:cytology* / Q000378:metabolism; D013997:Time Factors
keywords: Dopaminergic neuron; GABA and glycine receptors; Human induced pluripotent cells; Neural differentiation; Neural induction; Neuronal progenitors; Patch clamp; Voltage-gated currents