title: A Novel Protocol to Differentiate Induced Pluripotent Stem Cells by Neuronal microRNAs to Provide a Suitable Cellular Model.
abstract: Neurodegenerative diseases are one of the most challenging subjects in medicine. Investigation of their underlying genetic or epigenetic factors is hampered by lack of suitable models. Patient-specific induced pluripotent stem cells (iPS cells) represent a valuable approach to provide a proper model for poorly understood mechanisms of neuronal diseases and the related drug screenings. miR-124 and miR-128 are the two brain-enriched miRNAs with different time-points of expression during neuronal development. Herein, we transduced human iPS cells with miR-124 and miR-128 harboring lentiviruses sequentially. The transduced plasmids contained GFP and puromycin antibiotic-resistant genes for easier selection and identification. Morphological assessment and immunocytochemistry (overexpressions of beta-tubulin and neuron-specific enolase) confirmed that induced hiPS cells by miR-124 and miR-128 represent similar characteristics to those of mature neurons. In addition, the upregulation of neuron-specific enolase, beta-tubulin, Map2, GFAP, and BDNF was detected by quantitative real-time PCR. In conclusion, it seems that our novel protocol remarks the combinatorial effect of miR-124 and miR-128 on neural differentiation in the absence of any extrinsic factor. Moreover, such cellular models could be used in personalized drug screening and applied for more effective therapies.
authors.affiliation: Department of Cellular Biology, Stem Cell Technology Research Center, Tehran, Iran.|Neurosciences Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Mehrak
authors.initials: M
authors.lastname: Zare
authors.affiliation: Department of Haematology, School of Medicine, Tarbiat Modares University, Tehran, Iran.
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: Department of Medical Nanotechnology, Faculty of Advanced Medical Science, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Abolfazl
authors.initials: A
authors.lastname: Akbarzadeh
authors.affiliation: Department of Biotechnology, College of Science, University of Tehran, Tehran, Iran.
authors.forename: Behnaz
authors.initials: B
authors.lastname: Bakhshandeh
authors.affiliation: Department of Cellular Biology, Stem Cell Technology Research Center, Tehran, Iran.
authors.forename: Seyed Hamid
authors.initials: SH
authors.lastname: Aghaee-Bakhtiari
authors.affiliation: Department of Clinical Biochemistry and Laboratory, Faculty of Medicine, Tabriz University of Medical Sciences, Tabriz, Iran.|Department of Clinical Biochemistry, Radiopharmacy Lab, Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Nosratollah
authors.initials: N
authors.lastname: Zarghami
mesh_terms: D002454:Cell Differentiation / Q000235:genetics; D004353:Drug Evaluation, Preclinical / Q000379:methods; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000502:physiology*; D035683:MicroRNAs / Q000008:administration & dosage* / Q000096:biosynthesis / Q000235:genetics; D058953:Neural Stem Cells / Q000166:cytology / Q000502:physiology*; D019636:Neurodegenerative Diseases / Q000188:drug therapy / Q000235:genetics* / Q000473:pathology*; D009474:Neurons / Q000166:cytology / Q000502:physiology*; D010957:Plasmids / Q000008:administration & dosage / Q000235:genetics; D014161:Transduction, Genetic / Q000379:methods
chemical_list: C509207:MIRN124 microRNA, human; C545189:MIRN128 microRNA, human; D035683:MicroRNAs
keywords: cellular model; iPS cell; miR-124; miR-128; neuronal differentiation