title: Ob-Day15-TC2
organism: Mus musculus
description: Protocols: cells were extracted using TRIsure (Bioline, BIO-38033) following manufacturer' recommendations The mouse bone marrow stromal cell line ST2 established from Whitlock-Witte type long-term bone marrow culture of BC8 mice (Ogawa et al., 1988) was used during all experiments. Cells were grown in Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco, Life Technologies, 32404014) supplemented with 10% fetal bovine serum (FBS) (Gibco, Life Technologies, 10270-106, lot #41F8430K) and 1% L-Glutamine (Lonza, BE17-605E) in a constant atmosphere of 37°C and 5 % CO2. For differentiation into adipocytes and osteoblasts, ST2 cells were seeded 4 days before differentiation (day-4), reached 100% confluency after 48 hours (day-2) and were further maintained for 48 hours post-confluency (day 0). Adipogenic differentiation was subsequently initiated on day 0 (D0) by adding differentiation medium I consisting of growth medium, 0.5 mM isobutylmethylxanthine (IBMX) (Sigma-Aldrich, I5879), 0.25 µM dexamethasone (DEXA) (Sigma-Aldrich, D4902) and 5 µg/mL insulin (Sigma-Aldrich, I9278). From day 2 (D2) on differentiation medium II consisting of growth medium, 500 nM rosiglitazone (RGZ) (Sigma-Aldrich, R2408) and 5 µg/mL insulin (Sigma-Aldrich, I9278) was added and replaced every 2 days until 15 days of differentiation. Osteoblastic differentiation was induced with growth medium supplemented with 100 ng/mL bone morphogenetic protein-4 (BMP-4) (PeproTech, 315-27). Same media was replaced every 2 days until 15 days of osteoblastogenesis. Cells were extracted using TRIsure (Bioline, BIO-38033) following manufacturer' recommendations TruSeq unstranded mRNA Library Prep kit (Illumina) was used to prepare libraries
attributes.tag: ENA-FIRST-PUBLIC
attributes.value: 2019-02-08T17:02:37Z
attributes.tag: ENA-LAST-UPDATE
attributes.value: 2018-05-30T17:07:03Z
attributes.tag: External Id
attributes.value: SAMEA4699896
attributes.tag: INSDC center name
attributes.value: Life Sciences Research Unit, University of Luxembourg
attributes.tag: INSDC first public
attributes.value: 2019-02-08T17:02:37Z
attributes.tag: INSDC last update
attributes.value: 2018-05-30T17:07:03Z
attributes.tag: INSDC status
attributes.value: public
attributes.tag: Submitter Id
attributes.value: E-MTAB-6840:Ob-Day15-TC2
attributes.tag: broker name
attributes.value: ArrayExpress
attributes.tag: cell line
attributes.value: ST2
attributes.tag: cell type
attributes.value: osteoblast
attributes.tag: common name
attributes.value: house mouse
attributes.tag: organism part
attributes.value: bone marrow
attributes.tag: progenitor cell type
attributes.value: stromal cell
attributes.tag: sample name
attributes.value: E-MTAB-6840:Ob-Day15-TC2
attributes.tag: scientific_name
attributes.value: Mus musculus