title: Reprogrammed mouse fibroblasts differentiate into cells of the cardiovascular and hematopoietic lineages.
abstract: Forced expression of the four transcription factors Oct4, Sox2, c-Myc, and Klf4 is sufficient to confer a pluripotent state upon the murine fibroblast genome, generating induced pluripotent stem (iPS) cells. Although the differentiation potential of these cells is thought to be equivalent to that of embryonic stem (ES) cells, it has not been rigorously determined. In this study, we sought to identify the capacity of iPS cells to differentiate into Flk1-positive progenitors and their mesodermal progeny, including cells of the cardiovascular and hematopoietic lineages. Immunostaining of tissues from iPS cell-derived chimeric mice demonstrated that iPS cells could contribute in vivo to cardiomyocytes, smooth muscle cells, endothelial cells, and hematopoietic cells. To compare the in vitro differentiation potential of murine ES and iPS cells, we either induced embryoid body (EB) formation of each cell type or cultured the cells on collagen type IV (ColIV), an extracellular matrix protein that had been reported to direct murine ES cell differentiation to mesodermal lineages. EB formation and exposure to ColIV both induced iPS cell differentiation into cells that expressed cardiovascular and hematopoietic markers. To determine whether ColIV-differentiated iPS cells contained a progenitor cell with cardiovascular and hematopoietic differentiation potential, Flk1-positive cells were isolated by magnetic cell sorting and exposed to specific differentiation conditions, which induced differentiation into functional cardiomyocytes, smooth muscle cells, endothelial cells, and hematopoietic cells. Our data demonstrate that murine iPS cells, like ES cells, can differentiate into cells of the cardiovascular and hematopoietic lineages and therefore may represent a valuable cell source for applications in regenerative medicine. Disclosure of potential conflicts of interest is found at the end of this article.
authors.affiliation: Department of Medicine and Physiology, Cardiovascular Research Laboratory, University of California Los Angeles School of Medicine, Los Angeles, California 90095-1760, USA.
authors.forename: Katja
authors.initials: K
authors.lastname: Schenke-Layland
authors.forename: Katrin E
authors.initials: KE
authors.lastname: Rhodes
authors.forename: Ekaterini
authors.initials: E
authors.lastname: Angelis
authors.forename: Yekaterina
authors.initials: Y
authors.lastname: Butylkova
authors.forename: Sepideh
authors.initials: S
authors.lastname: Heydarkhan-Hagvall
authors.forename: Christos
authors.initials: C
authors.lastname: Gekas
authors.forename: Rui
authors.initials: R
authors.lastname: Zhang
authors.forename: Joshua I
authors.initials: JI
authors.lastname: Goldhaber
authors.forename: Hanna K
authors.initials: HK
authors.lastname: Mikkola
authors.forename: Kathrin
authors.initials: K
authors.lastname: Plath
authors.forename: W Robb
authors.initials: WR
authors.lastname: MacLellan
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000502:physiology*; D004730:Endothelium, Vascular / Q000166:cytology*; D005347:Fibroblasts / Q000166:cytology*; D017930:Genes, Reporter; D016678:Genome; D049452:Green Fluorescent Proteins / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D000090062:Kruppel-Like Factor 4; D051379:Mice; D009130:Muscle, Smooth / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology*; D014157:Transcription Factors / Q000235:genetics / Q000502:physiology
chemical_list: C000715468:Klf4 protein, mouse; D000090062:Kruppel-Like Factor 4; D014157:Transcription Factors; D049452:Green Fluorescent Proteins