View source for MSC to adipocyte (mouse)

{{TimeCourse
|TCOverview=Senile osteoporosis is the most common metabolic bone disease. This disease is often accompanied by increasing adipocytes in bone marrow tissues [1]. The ectopic adipocytes differentiation following bone loss seems to be caused by unbalanced differentiation of mesenchymal stem cells (MSCs) [2]. Although several differentiation regulators of MSCs have already been reported, little is known about the regulatory dynamics of bi-directional adipocytes/osteoblasts differentiation.<br>To uncover the complex mechanism of osteoporosis and metabolic disease, we performed a variety of genome-wide analyses about gene expression and regulation that could influence adipocytes/osteoblasts differentiation from mouse ST2 cells (bone marrow-derived stromal cell line) [3,4]. In particular, we focused on the differentiation-specific non-cording RNAs and antisense transcripts as the novel regulator candidates of adipocytes/osteoblasts differentiation.<br><br>References:<br>1. Burkhardt R, Kettner G, Bohm W, Schmidmeier M, Schlag R, et al. Changes in trabecular bone, hematopoiesis and bone marrow vessels in aplastic anemia, primary osteoporosis, and old age: a comparative histomorphometric study. Bone (1987) 8(3):157-164. <html><a href='https://www.ncbi.nlm.nih.gov/pubmed/3606907' target='_blank'>PMID:3606907</a></html><br>2. Nuttall ME, Gimble JM. Controlling the balance between osteoblastogenesis and adipogenesis and the consequent therapeutic implications. Curr Opin Pharmacol (2004) 4(3):290–294. <html><a href='https://www.ncbi.nlm.nih.gov/pubmed/15140422' target='_blank'>PMID:15140422</a></html><br>3. Tokuzawa Y, Yagi K, Yamashita Y, Nakachi Y, Nikaido I, et al. Id4, a new candidate gene for senile osteoporosis, acts as a molecular switch promoting osteoblast differentiation. PLoS Genet (2010) 6(7):e1001019. <html><a href='https://www.ncbi.nlm.nih.gov/pubmed/20628571' target='_blank'>PMID:20628571</a></html><br>4. Mizuno Y, Yagi K, Tokuzawa Y, Kanesaki-Yatsuka Y, Suda T, et al. miR-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation. Biochem Biophys Res Commun (2008) 368(2):267-272. <html><a href='https://www.ncbi.nlm.nih.gov/pubmed/18230348' target='_blank'>PMID:18230348</a></html>.<br>
|TCQuality_control='''Marker gene expression:'''<br>Adipocyte differentation, hCAGE:<br><html><img src='/resource_browser/images/TC_qc/Adipo1.jpg'><img src='/resource_browser/images/TC_qc/Adipo2.jpg'><img src='/resource_browser/images/TC_qc/Legend.jpg'><br><img src='/resource_browser/images/TC_qc/Adipo3.jpg'><img src='/resource_browser/images/TC_qc/Adipo4.jpg'><img src='/resource_browser/images/TC_qc/Legend.jpg'><br><img src='/resource_browser/images/TC_qc/Adipo5.jpg'><img src='/resource_browser/images/TC_qc/Adipo6.jpg'><img src='/resource_browser/images/TC_qc/Legend.jpg'><br><img src='/resource_browser/images/TC_qc/Osteo1.jpg'><img src='/resource_browser/images/TC_qc/Osteo2.jpg'><img src='/resource_browser/images/TC_qc/Legend.jpg'><br><img src='/resource_browser/images/TC_qc/Osteo3.jpg'><img src='/resource_browser/images/TC_qc/Osteo4.jpg'><img src='/resource_browser/images/TC_qc/Legend.jpg'><br><img src='/resource_browser/images/TC_qc/Osteo5.jpg'><img src='/resource_browser/images/TC_qc/Osteo6.jpg'><img src='/resource_browser/images/TC_qc/Legend.jpg'><br></html><br>Figurea 4. Gene expression (TPM) of key regulators (Hes1, Id1, Id2,Sp7) and differentiation markers (Alpl, Bglap).<br>
|TCSample_description='''Cell line:'''<br>ST2 cells were obtained from RIKEN BioResource Center (BRC, Tsukuba, Japan). These cell line is bone marrow-derived stromal cell line. ST2 differentiated most efficiently into both osteoblasts and adipocytes [3].<br>'''Cell culture:'''<br>ST2 cells were cultured according to the protocols supplied by BRC (RPMI1640 supplemented with 10% fetal bovine serum) [4].<br>'''Differentiation induction:'''<br>''Adipocyte differentation:''<br>Adipogenic differentiation was induced by changing the medium to differentiation medium supplemented with 10% fetal bovine serum (FBS), 0.5 mM 3-isobutyl-1-methlxanthine, 0.25 mM dexamethasone, and insulin-transferrin-selenium-X supplement containing 5 mg/ml of insulin (Invitrogen, Carlsbad, CA) and 1 mM rosiglitazone. After 48 hr, the differentiation medium was replaced with conditional culture medium supplemented with 10% FBS [3]. Pre-conditioned medium which was originally same as culure medium, was carried by parallel culturing of ST2, to avoid the perturbation by "medium-change" shock as soon as possible.<br><html><img src='/resource_browser/images/TC_qc/NileRed4d.jpg'></html><br>Figure 1. Histological staining of ST2 cells using Nile Red staining during adipocyte differentiation.<br>The number of Nile Red stained lipid droplets increased in ST2 cell (4 days after adipocyte induction). Bar: 100 μm.<br><br>''Osteoblast differentiation:''<br>Osteogenic differentiation was induced by changing the medium every three days to culture medium supplemented with 100 ng/ml of bone morphogenetic protein 4 (BMP4, R&D Systems, Mineapolis, MN) [3]. Pre-conditioned medium which was originally same as culure medium, was carried by parallel culturing of ST2, to avoid the perturbation by "medium-change" shock as soon as possible.<br><html><img src='/resource_browser/images/TC_qc/ALP20d.jpg'><img src='/resource_browser/images/TC_qc/ALP6d.jpg'><img src='/resource_browser/images/TC_qc/ALP0d.jpg'></html><br>Figure 2. ALP staining of ST2 cells for 0, 6 and 20 days after osteoblast induction.<br>The ALP activity of ST2 cells (right, 20 days) were more prominently increased in the presence of BMP4 than 0 day (left).<br><br>'''Sampling:'''<br>ST2 cells are sampled during adipocyte or osteoblast differentiation (15min, 30min, 1-3hr, 6,12,18,24,36,48hr, 3-6day) and non-treatment control (2 points; 0,6day).<br><html><img src='/resource_browser/images/TC_qc/500px-Figure1.png'></html><br>Figure 3. Sampling points for ST2 time-course CAGE data<br><br>References:<br>3. Tokuzawa Y, Yagi K, Yamashita Y, Nakachi Y, Nikaido I, et al. Id4, a new candidate gene for senile osteoporosis, acts as a molecular switch promoting osteoblast differentiation. PLoS Genet (2010) 6(7):e1001019. <html><a href='https://www.ncbi.nlm.nih.gov/pubmed/20628571' target='_blank'>PMID:20628571</a></html><br>4. Mizuno Y, Yagi K, Tokuzawa Y, Kanesaki-Yatsuka Y, Suda T, et al. miR-125b inhibits osteoblastic differentiation by down-regulation of cell proliferation. Biochem Biophys Res Commun (2008) 368(2):267-272. <html><a href='https://www.ncbi.nlm.nih.gov/pubmed/18230348' target='_blank'>PMID:18230348</a></html>.<br>
|Time_Course=
|category_treatment=Differentiation
|collaborators=Yasushi Okazaki
|description=mouse_ST2_adipocytes
|germ_layer=mesoderm
|libraryids=CNhs13114,CNhs13115,CNhs13116,CNhs13117,CNhs13118,CNhs13120,CNhs13121,CNhs13122,CNhs13123,CNhs13125,CNhs13126,CNhs13127,CNhs13128,CNhs13129,CNhs13243,CNhs13244,CNhs13245,CNhs13247,CNhs13248,CNhs13249,CNhs13250,CNhs13251,CNhs13252,CNhs13253,CNhs13254,CNhs13255,CNhs13256,CNhs13257,CNhs13258,CNhs13292,CNhs13293,CNhs13295,CNhs13296,CNhs13297,CNhs13298,CNhs13299,CNhs13300,CNhs13301,CNhs13302,CNhs13303,CNhs13304,CNhs13305,CNhs13306,CNhs13307
|number_time_points=16
|page_name=mouse_MSC_adipocytes
|primary_cells=primary cells
|series=IN_VITRO DIFFERENTIATION SERIES
|species=Mouse (Mus musculus)
|tet_config=https://fantom.gsc.riken.jp/5/suppl/tet/MSC_adipocytes.tsv.gz
|tet_file=https://fantom.gsc.riken.jp/5/tet#!/search/?filename=mm9.cage_peak_phase1and2combined_tpm_ann_decoded.osc.txt.gz&file=1&c=1&c=260&c=261&c=262&c=263&c=264&c=265&c=266&c=267&c=268&c=269&c=270&c=271&c=272&c=273&c=274&c=275&c=276&c=277&c=278&c=279&c=280&c=281&c=282&c=283&c=284&c=285&c=286&c=288&c=289&c=290&c=291&c=292&c=293&c=294&c=295&c=296&c=297&c=298&c=299&c=300&c=301&c=302&c=303&c=304&c=350&c=351&c=352&c=353&c=354&c=355
|time_points=
|time_span=6 days
|timepoint_design=Early focus
|tissue_cell_type=Mesenchymal>>adipose
|zenbu_config=https://fantom.gsc.riken.jp/zenbu/gLyphs/#config=af0apQV7MN6nLTx_ohbbPB
}}