title: Neural differentiation of patient specific iPS cells as a novel approach to study the pathophysiology of multiple sclerosis.
abstract: The recent introduction of technologies capable of reprogramming human somatic cells into induced pluripotent stem (iPS) cells offers a unique opportunity to study many aspects of neurodegenerative diseases in vitro that could ultimately lead to novel drug development and testing. Here, we report for the first time that human dermal fibroblasts from a patient with relapsing-remitting Multiple Sclerosis (MS) were reprogrammed to pluripotency by retroviral transduction using defined factors (OCT4, SOX2, KLF4, and c-MYC). The MSiPS cell lines resembled human embryonic stem (hES) cell-like colonies in morphology and gene expression and exhibited silencing of the retroviral transgenes after four passages. MSiPS cells formed embryoid bodies that expressed markers of all three germ layers by immunostaining and Reverse Transcriptase (RT)-PCR. The injection of undifferentiated iPS cell colonies into immunodeficient mice formed teratomas, thereby demonstrating pluripotency. The MSiPS cells were successfully differentiated into mature astrocytes, oligodendrocytes and neurons with normal karyotypes. Although MSiPS-derived neurons displayed some differences in their electrophysiological characteristics as compared to the control cell line, they exhibit properties of functional neurons, with robust resting membrane potentials, large fast tetrodotoxin-sensitive action potentials and voltage-gated sodium currents. This study provides for the first time proof of concept that disease cell lines derived from skin cells obtained from an MS patient can be generated and successfully differentiated into mature neural lineages. This represents an important step in a novel approach for the study of MS pathophysiology and potential drug discovery.
authors.affiliation: Monash Immunology and Stem Cell Laboratories, Monash University, Wellington Road, Clayton, Victoria 3800, Australia.
authors.forename: Bi
authors.initials: B
authors.lastname: Song
authors.forename: Guizhi
authors.initials: G
authors.lastname: Sun
authors.forename: Daniella
authors.initials: D
authors.lastname: Herszfeld
authors.forename: Aude
authors.initials: A
authors.lastname: Sylvain
authors.forename: Naomi V
authors.initials: NV
authors.lastname: Campanale
authors.forename: Claire E
authors.initials: CE
authors.lastname: Hirst
authors.forename: Sally
authors.initials: S
authors.lastname: Caine
authors.forename: Helena C
authors.initials: HC
authors.lastname: Parkington
authors.forename: Mary A
authors.initials: MA
authors.lastname: Tonta
authors.forename: Harold A
authors.initials: HA
authors.lastname: Coleman
authors.forename: Martin
authors.initials: M
authors.lastname: Short
authors.forename: Sharon D
authors.initials: SD
authors.lastname: Ricardo
authors.forename: Benjamin
authors.initials: B
authors.lastname: Reubinoff
authors.forename: Claude C A
authors.initials: CC
authors.lastname: Bernard
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D019070:Cell Lineage; D055724:Electrophysiological Phenomena; D005347:Fibroblasts / Q000473:pathology; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000473:pathology*; D000090062:Kruppel-Like Factor 4; D051379:Mice; D016513:Mice, SCID; D018895:Microsatellite Repeats / Q000235:genetics; D009103:Multiple Sclerosis / Q000473:pathology* / Q000503:physiopathology*; D009474:Neurons / Q000473:pathology*; D050814:Octamer Transcription Factor-3 / Q000235:genetics; D009836:Oligodendroglia / Q000473:pathology; D039904:Pluripotent Stem Cells / Q000473:pathology; D011401:Promoter Regions, Genetic / Q000235:genetics; D012190:Retroviridae / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000473:pathology; D014161:Transduction, Genetic
chemical_list: C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D050814:Octamer Transcription Factor-3