title: DOR2AO
organism: Homo sapiens
description: Protocols: Bone-marrow human mesenchymal stromal-cells were obtained from three donors recruited in the Bone-Marrow Transplant Program of Hematology Service at Reina Sofía University Hospital (Córdoba, Spain), after signed written informed-consent. MSC were cultured in Minimum Essential-Medium Alpha (MEM α) Eagle from Lonza (Basel, Switzerland), supplemented with 10% fetal bovine-serum (FBS) from Gibco - Life Technologies - Thermo Fisher Scientific (Waltham, MA, USA), 2 mM UltraGlutamine (Lonza), 1 ng basic fibroblast-growth factor (bFGF)/ml from Sigma-Aldrich (Saint Louis, MO, USA), 100 U ampicillin and 0.1 mg streptomycin/ml. Cells were grown at 37 ºC and 5% CO2 in humidity. Cultures grown until about 80 to 90% confluence were maintained in MEMα plus 10% FBS without bFGF (controls, C) or likewise but supplemented with 5x10-7 M dexamethasone, 0.5 mM isobutylmethylxanthine and 50 mM indomethacin (Sigma-Aldrich) adipogenic inductors for cell differentiation, in absence (A) or presence (A+O) 10-5 M oleuropein (purity >80%) from Extrasynthese (Lyon, France). Adipogenic differentiation at day 14 was determined by oil-red O staining of lipid droplets. In short, cells were fixed with 3.7% formaldehyde, washed with 60% isopropanol in water and stained with a mixture of 60% of 0.3% (w/v in isopropanol) of oil-red O and 40% distilled water for 20 minutes. They were then washed with water, stained with hematoxylin (all chemicals from Sigma-Aldrich) and visualized under a light microscope. Total RNA was isolated at days seven and 14 after adipogenesis induction, using mirVana miRNA Isolation Kit from Ambion - Life Technologies - Thermo Fisher Scientific. RNA were quantified using a ND-1000 spectrophotometer from NanoDrop - Thermo Fisher Scientific. Integrity was checked by agarose-gel electrophoresis (AGE). RNA samples were pooled from each time and donor, to generate three samples: control (C), adipocytes (A) and adipocytes plus oleuropein (A+O) for gene-expression analyses.
attributes.tag: ENA first public
attributes.value: 2019-01-09
attributes.tag: ENA last update
attributes.value: 2016-11-09
attributes.tag: External Id
attributes.value: SAMEA4537140
attributes.tag: INSDC center alias
attributes.value: Unidad de Metabolismo Mineral, Instituto Maimonides de Investigacion Biomedica de Cordoba (IMIBIC), Hospital Universitario Reina Sofia, Avda. Menendez Pidal s/n, 14004 Cordoba,
attributes.tag: INSDC center name
attributes.value: Unidad de Metabolismo Mineral, Instituto Maimonides de Investigacion Biomedica de Cordoba (IMIBIC), Hospital Universitario Reina Sofia, Avda. Menendez Pidal s/n, 14004 Cordoba,
attributes.tag: INSDC first public
attributes.value: 2019-01-09T17:03:19Z
attributes.tag: INSDC last update
attributes.value: 2016-11-09T09:53:39Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-5211:DOR2AO
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell type
attributes.value: adipocyte
attributes.tag: common name
attributes.value: human
attributes.tag: organism part
attributes.value: bone marrow
attributes.tag: progenitor cell type
attributes.value: mesenchymal stem cell
attributes.tag: sample name
attributes.value: E-MTAB-5211:DOR2AO