MSC to adipocyte (mouse): Difference between revisions
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|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. < | |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. <span class="plainlinks">[https://www.ncbi.nlm.nih.gov/pubmed/3606907 PMID:3606907]</span><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. <span class="plainlinks">[https://www.ncbi.nlm.nih.gov/pubmed/15140422 PMID:15140422]</span><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. <span class="plainlinks">[https://www.ncbi.nlm.nih.gov/pubmed/20628571 PMID:20628571]</span><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. <span class="plainlinks">[https://www.ncbi.nlm.nih.gov/pubmed/18230348 PMID:18230348]</span>.<br> | ||
|TCQuality_control='''Marker gene expression:'''<br>Adipocyte differentation, hCAGE:<br><html><img src=' | |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=' | |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. <span class="plainlinks">[https://www.ncbi.nlm.nih.gov/pubmed/20628571 PMID:20628571]</span><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. <span class="plainlinks">[https://www.ncbi.nlm.nih.gov/pubmed/18230348 PMID:18230348]</span>.<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 14: | Line 14: | ||
|series=IN_VITRO DIFFERENTIATION SERIES | |series=IN_VITRO DIFFERENTIATION SERIES | ||
|species=Mouse (Mus musculus) | |species=Mouse (Mus musculus) | ||
|tet_config= | |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_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= | |zenbu_config=https://fantom.gsc.riken.jp/zenbu/gLyphs/#config=af0apQV7MN6nLTx_ohbbPB | ||
}} | }} | ||
Latest revision as of 18:01, 14 March 2022
| Series: | IN_VITRO DIFFERENTIATION SERIES |
|---|---|
| Species: | Mouse (Mus musculus) |
| Genomic View: | Zenbu |
| Expression table: | FILE |
| Link to TET: | TET |
| Sample providers : | Yasushi Okazaki |
| Germ layer: | mesoderm |
| Primary cells or cell line: | primary cells |
| Time span: | 6 days |
| Number of time points: | 16 |
| Overview |
|---|
|
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. |
| Sample description |
|---|
|
Cell line: |
| Quality control |
|---|
|
Marker gene expression: |
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
| 12308-130F4 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 00hr15min | biol_rep1 (015mA1) |
| 12309-130F5 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 00hr30min | biol_rep1 (030mA1) |
| 12310-130F6 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 01hr | biol_rep1 (001hA1) |
| 12311-130F7 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 02hr | biol_rep1 (002hA1) |
| 12312-130F8 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 03hr | biol_rep1 (003hA1) |
| 12313-130F9 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 06hr | biol_rep1 (006hA1) |
| 12314-130G1 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 12hr | biol_rep1 (012hA1) |
| 12315-130G2 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 18hr | biol_rep1 (018hA1) |
| 12316-130G3 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 24hr | biol_rep1 (024hA1) |
| 12318-130G5 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day02 | biol_rep1 (048hA1) |
| 12319-130G6 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day03 | biol_rep1 (072hA1) |
| 12320-130G7 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day04 | biol_rep1 (096hA1) |
| 12321-130G8 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day05 | biol_rep1 (120hA1) |
| 12322-130G9 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day06 | biol_rep1 (144hA1) |
| 12338-130I7 | ST2 (Mesenchymal stem cells) cells, medium change (without induction) | 00hr | biol_rep1 |
| 12347-131A7 | ST2 (Mesenchymal stem cells) cells, medium change (without induction) | day06 | biol_rep1 |
| 12430-132A9 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 00hr15min | biol_rep2 (015mA2) |
| 12431-132B1 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 00hr30min | biol_rep2 (030mA2) |
| 12432-132B2 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 01hr | biol_rep2 (001hA2) |
| 12433-132B3 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 02hr | biol_rep2 (002hA2) |
| 12434-132B4 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 03hr | biol_rep2 (003hA2) |
| 12435-132B5 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 06hr | biol_rep2 (006hA2) |
| 12436-132B6 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 12hr | biol_rep2 (012hA2) |
| 12437-132B7 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 18hr | biol_rep2 (018hA2) |
| 12438-132B8 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 24hr | biol_rep2 (024hA2) |
| 12439-132B9 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 36hr | biol_rep2 (036hA2) |
| 12440-132C1 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day02 | biol_rep2 (048hA2) |
| 12441-132C2 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day03 | biol_rep2 (072hA2) |
| 12442-132C3 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day04 | biol_rep2 (096hA2) |
| 12443-132C4 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day05 | biol_rep2 (120hA2) |
| 12444-132C5 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day06 | biol_rep2 (144hA2) |
| 12460-132E3 | ST2 (Mesenchymal stem cells) cells, medium change (without induction) | 00hr | biol_rep2 |
| 12469-132F3 | ST2 (Mesenchymal stem cells) cells, medium change (without induction) | day06 | biol_rep2 |
| 12552-133F5 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 00hr15min | biol_rep3 (015mA3) |
| 12553-133F6 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 00hr30min | biol_rep3 (030mA3) |
| 12554-133F7 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 01hr | biol_rep3 (001hA3) |
| 12555-133F8 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 02hr | biol_rep3 (002hA3) |
| 12556-133F9 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 03hr | biol_rep3 (003hA3) |
| 12557-133G1 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 06hr | biol_rep3 (006hA3) |
| 12558-133G2 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 12hr | biol_rep3 (012hA3) |
| 12559-133G3 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 18hr | biol_rep3 (018hA3) |
| 12560-133G4 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 24hr | biol_rep3 (024hA3) |
| 12561-133G5 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | 36hr | biol_rep3 (036hA3) |
| 12562-133G6 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day02 | biol_rep3 (048hA3) |
| 12563-133G7 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day03 | biol_rep3 (072hA3) |
| 12564-133G8 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day04 | biol_rep3 (096hA3) |
| 12565-133G9 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day05 | biol_rep3 (120hA3) |
| 12566-133H1 | ST2 (Mesenchymal stem cells) cells, differentiation to adipocytes | day06 | biol_rep3 (144hA3) |
| 12582-133I8 | ST2 (Mesenchymal stem cells) cells, medium change (without induction) | 00hr | biol_rep3 |
| 12591-134A8 | ST2 (Mesenchymal stem cells) cells, medium change (without induction) | day06 | biol_rep3 |
