title: Directing mouse embryonic neurosphere differentiation toward an enriched neuronal population.
abstract: Neural stem cells (NSC) are self-renewing multipotent cells that have emerged as a powerful tool to repair the injured brain. These cells can be cultured as neurospheres, which are floating aggregates of neural stem/progenitor cells (NSPCs). Despite their high clonal expansion capacity, it has been suggested that in neurospheres, only a small percentage of cells are capable of proliferation and that this system is not efficient in terms of neurogenic competence. Thus, our aim was to develop a neurosphere culture method with a highly proliferative stem/progenitor cell population and particularly with a prominent neurogenic potential, surpassing some of the claimed weaknesses of the neurosphere assay. In our model, mouse neurospheres were harvested from neural tissue at E15 and after only 4 days in vitro (DIV), we have achieved highly proliferative primary neurospheres (81% Sox2 and 76% Ki67 positive cells) and a rather low number of cells expressing glial and neuronal markers (∼10%). After inducing differentiation, we have attained an enriched neuronal population (45% β-III-tubulin positive cells at 15 DIV). Using a simple methodology, we have developed a NSPC model that can provide a valuable source of neuronal precursors, thus offering a potential starting point for cell replacement therapies following CNS injury.
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal.
authors.forename: Ema F
authors.initials: EF
authors.lastname: Torrado
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal.
authors.forename: Cátia
authors.initials: C
authors.lastname: Gomes
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal.
authors.forename: Gisela
authors.initials: G
authors.lastname: Santos
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal; Department of Biochemistry and Human Biology, Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal.
authors.forename: Adelaide
authors.initials: A
authors.lastname: Fernandes
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal; Department of Biochemistry and Human Biology, Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal. Electronic address: dbrites@ff.ul.pt.
authors.forename: Dora
authors.initials: D
authors.lastname: Brites
authors.affiliation: Research Institute for Medicines (iMed.ULisboa), Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal; Department of Biochemistry and Human Biology, Faculdade de Farmácia, Universidade de Lisboa, Avenida Professor Gama Pinto, 1649-003 Lisbon, Portugal. Electronic address: asfalcao@ff.ul.pt.
authors.forename: Ana S
authors.initials: AS
authors.lastname: Falcão
mesh_terms: D000818:Animals; D001921:Brain / Q000166:cytology* / Q000196:embryology; D002454:Cell Differentiation / Q000502:physiology*; D049109:Cell Proliferation; D002478:Cells, Cultured; D004622:Embryo, Mammalian; D005260:Female; D019394:Ki-67 Antigen / Q000378:metabolism; D051379:Mice; D058953:Neural Stem Cells / Q000502:physiology*; D009457:Neuroglia; D009474:Neurons / Q000502:physiology*; D011247:Pregnancy; D055748:SOXB1 Transcription Factors / Q000378:metabolism; D013997:Time Factors
chemical_list: D019394:Ki-67 Antigen; D055748:SOXB1 Transcription Factors; C527784:Sox2 protein, rat
keywords: Mouse cortex E15 neurospheres; Mouse neural stem cells; Neural progenitor cells; Neuronal differentiation