title: Human induced pluripotent stem cells derived from fetal neural stem cells successfully undergo directed differentiation into cartilage.
abstract: Induced pluripotent stem (iPS) cells can be derived from a wide range of somatic cells via overexpression of a set of specific genes. With respect to their properties, iPS cells closely resemble embryonic stem cells. Because of their main property, pluripotency, iPS cells have excellent prospects for use in substitutive cell therapy; however, the methods of directed differentiation of iPS cells have not been yet sufficiently elaborated. In this work, we derived human iPS cells from fetal neural stem (FNS) cells by transfection with a polycistronic plasmid vector carrying the mouse Oct4, Sox2, Klf4, and c-Myc genes or a plasmid expressing the human OCT4 gene. We have shown that human FNS cells can be effectively reprogrammed despite a low transfection level (10%-15%) and that the use of 2-propylvaleric (valproic) acid and BIX-01294 increases the yield of iPS cell clones to ∼7-fold. Further, transient expression of OCT4 alone is sufficient for reprogramming. The iPS cells obtained express all the major markers of embryonic stem cells and are able to differentiate in vitro into ectodermal, mesodermal, and endodermal derivatives. In addition, we have found that the human iPS cells derived from FNS cells can be successfully subjected to in vitro directed chondrogenic differentiation to form functional cartilaginous tissue.
authors.affiliation: Institute of Cytology and Genetics, Russian Academy of Sciences, Novosibirsk, Russia.
authors.forename: Sergey P
authors.initials: SP
authors.lastname: Medvedev
authors.forename: Elena V
authors.initials: EV
authors.lastname: Grigor'eva
authors.forename: Alexander I
authors.initials: AI
authors.lastname: Shevchenko
authors.forename: Anastasia A
authors.initials: AA
authors.lastname: Malakhova
authors.forename: Elena V
authors.initials: EV
authors.lastname: Dementyeva
authors.forename: Alexander A
authors.initials: AA
authors.lastname: Shilov
authors.forename: Evgeny A
authors.initials: EA
authors.lastname: Pokushalov
authors.forename: Alla M
authors.initials: AM
authors.lastname: Zaidman
authors.forename: Maria A
authors.initials: MA
authors.lastname: Aleksandrova
authors.forename: Egor Yu
authors.initials: EY
authors.lastname: Plotnikov
authors.forename: Gennady T
authors.initials: GT
authors.lastname: Sukhikh
authors.forename: Suren M
authors.initials: SM
authors.lastname: Zakian
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002356:Cartilage / Q000166:cytology* / Q000378:metabolism; D002452:Cell Count; D002454:Cell Differentiation / Q000235:genetics; D048430:Cell Shape; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D053686:Fetal Stem Cells / Q000166:cytology* / Q000378:metabolism; D005786:Gene Expression Regulation; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000201:enzymology; D000090062:Kruppel-Like Factor 4; D051379:Mice; D018895:Microsatellite Repeats / Q000235:genetics; D058953:Neural Stem Cells / Q000166:cytology* / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
chemical_list: D015415:Biomarkers; C000715467:KLF4 protein, human; C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D000469:Alkaline Phosphatase