MSC to adipocyte (mouse): Difference between revisions
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|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='https://fantom5-collaboration.gsc.riken.jp/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='https://fantom5-collaboration.gsc.riken.jp/resource_browser/images/TC_qc/ALP20d.jpg'><img src='https://fantom5-collaboration.gsc.riken.jp/resource_browser/images/TC_qc/ALP6d.jpg'><img src='https://fantom5-collaboration.gsc.riken.jp/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='https://fantom5-collaboration.gsc.riken.jp/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='http://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='http://www.ncbi.nlm.nih.gov/pubmed/18230348' target='_blank'>PMID:18230348</a></html>.<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='https://fantom5-collaboration.gsc.riken.jp/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='https://fantom5-collaboration.gsc.riken.jp/resource_browser/images/TC_qc/ALP20d.jpg'><img src='https://fantom5-collaboration.gsc.riken.jp/resource_browser/images/TC_qc/ALP6d.jpg'><img src='https://fantom5-collaboration.gsc.riken.jp/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='https://fantom5-collaboration.gsc.riken.jp/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='http://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='http://www.ncbi.nlm.nih.gov/pubmed/18230348' target='_blank'>PMID:18230348</a></html>.<br> | ||
|Time_Course= | |Time_Course= | ||
|category_treatment= | |category_treatment=Differentiation | ||
|collaborators=Yasushi Okazaki | |collaborators=Yasushi Okazaki | ||
|description=mouse_ST2_adipocytes | |description=mouse_ST2_adipocytes | ||
Line 17: | Line 17: | ||
|time_points= | |time_points= | ||
|time_span=6 days | |time_span=6 days | ||
|timepoint_design= | |timepoint_design=Early focus | ||
|tissue_cell_type= | |tissue_cell_type=Mesenchymal>>adipose | ||
|zenbu_config=http://fantom.gsc.riken.jp/zenbu/gLyphs/#config=eDHhE-APFBJsq9d4oP_U7 | |zenbu_config=http://fantom.gsc.riken.jp/zenbu/gLyphs/#config=eDHhE-APFBJsq9d4oP_U7 | ||
}} | }} |
Revision as of 12:31, 4 February 2015
Series: | IN_VITRO DIFFERENTIATION SERIES |
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Species: | Mouse (Mus musculus) |
Genomic View: | Zenbu |
Expression table: | FILE |
Link to TET: | [{{{tet_file}}} TET] |
Sample providers : | Yasushi Okazaki |
Germ layer: | mesoderm |
Primary cells or cell line: | primary cells |
Time span: | 6 days |
Number of time points: | 16 |
CollapseOverview |
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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. |
ExpandSample description |
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ExpandQuality control |
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Profiled time course samples
Only samples that passed quality controls (Arner et al. 2015) are shown here. The entire set of samples are downloadable from FANTOM5 human / mouse samples