title: Ultrastructural study of mouse adipose-derived stromal cells induced towards osteogenic direction.
abstract: We investigated the ultrastructural characteristics of mouse adipose-derived stem/stromal cells (ASCs) induced towards osteogenic lineage. ASCs were isolated from adipose tissue of FVB-Cg-Tg(GFPU)5Nagy/J mice and expanded in monolayer culture. Flow cytometry, histochemical staining, and electron microscopy techniques were used to characterize the ASCs with respect to their ability for osteogenic differentiation capacity. Immunophenotypically, ASCs were characterized by high expression of the CD44 and CD90 markers, while the relative content of cells expressing CD45, CD34 and CD117 markers was <2%. In assays of differentiation, the positive response to osteogenic differentiation factors was observed and characterized by deposition of calcium in the extracellular matrix and alkaline phosphatase production. Electron microscopy analysis revealed that undifferentiated ASCs had a rough endoplasmic reticulum with dilated cisterns and elongated mitochondria. At the end of the osteogenic differentiation, the ASCs transformed from their original fibroblast-like appearance to having a polygonal osteoblast-like morphology. Ultrastructurally, these cells were characterized by large euchromatic nucleus and numerous cytoplasm containing elongated mitochondria, a very prominent rough endoplasmic reticulum, Golgi apparatus and intermediate filament bundles. Extracellular matrix vesicles of variable size similar to the calcification nodules were observed among collagen fibrils. Our data provide the ultrastructural basis for further studies on the cellular mechanisms involved in osteogenic differentiation of mouse adipose-derived stem/stromal cells. Microsc. Res. Tech. 79:557-564, 2016. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Cytology, Bogomoletz Institute of Physiology, Kyiv, Ukraine.|Cell and Tissue Technologies Department, State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine.
authors.forename: Oleg
authors.initials: O
authors.lastname: Tsupykov
authors.affiliation: Cell and Tissue Technologies Department, State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine.
authors.forename: Alina
authors.initials: A
authors.lastname: Ustymenko
authors.affiliation: Cell and Tissue Technologies Department, State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine.
authors.forename: Vitaliy
authors.initials: V
authors.lastname: Kyryk
authors.affiliation: Department of Cytology, Bogomoletz Institute of Physiology, Kyiv, Ukraine.
authors.forename: Ekaterina
authors.initials: E
authors.lastname: Smozhanik
authors.affiliation: Department of Cytology, Bogomoletz Institute of Physiology, Kyiv, Ukraine.
authors.forename: Kateryna
authors.initials: K
authors.lastname: Yatsenko
authors.affiliation: Cell and Tissue Technologies Department, State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine.
authors.forename: Gennadii
authors.initials: G
authors.lastname: Butenko
authors.affiliation: Department of Cytology, Bogomoletz Institute of Physiology, Kyiv, Ukraine.|Cell and Tissue Technologies Department, State Institute of Genetic and Regenerative Medicine, Kyiv, Ukraine.
authors.forename: Galina
authors.initials: G
authors.lastname: Skibo
mesh_terms: D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D015703:Antigens, CD; D002454:Cell Differentiation / Q000502:physiology*; D002478:Cells, Cultured; D005434:Flow Cytometry; D016130:Immunophenotyping; D051379:Mice; D046529:Microscopy, Electron, Transmission; D010012:Osteogenesis / Q000502:physiology*; D017154:Stromal Cells / Q000737:chemistry / Q000166:cytology / Q000648:ultrastructure*
chemical_list: D015703:Antigens, CD
keywords: adipose-derived stem/stromal cells; directed osteogenic differentiation; electron microscopy