MSC to osteocyte (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_osteocytes | |description=mouse_ST2_osteocytes | ||
| 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_osteocytes.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=350&c=351&c=352&c=353&c=354&c=355&c=305&c=306&c=307&c=308&c=309&c=310&c=311&c=312&c=313&c=314&c=315&c=316&c=317&c=318&c=319&c=320&c=321&c=322&c=323&c=324&c=325&c=326&c=327&c=328&c=329&c=330&c=331&c=332&c=333&c=334&c=335&c=336&c=337&c=338&c=339&c=340&c=341&c=342&c=343&c=344&c=345&c=346&c=347&c=348&c=349 | |||
|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>>ostecyte | ||
|zenbu_config= | |zenbu_config=https://fantom.gsc.riken.jp/zenbu/gLyphs/#config=HVSblaPHMZvo-UNxMlnbQD | ||
}} | }} | ||
Latest revision as of 17:55, 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
| 12323-130H1 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 00hr15min | biol_rep1 (015mB1) |
| 12324-130H2 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 00hr30min | biol_rep1 (030mB1) |
| 12325-130H3 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 01hr | biol_rep1 (001hB1) |
| 12326-130H4 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 02hr | biol_rep1 (002hB1) |
| 12327-130H5 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 03hr | biol_rep1 (003hB1) |
| 12328-130H6 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 06hr | biol_rep1 (006hB1) |
| 12329-130H7 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 12hr | biol_rep1 (012hB1) |
| 12330-130H8 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 18hr | biol_rep1 (018hB1) |
| 12331-130H9 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 24hr | biol_rep1 (024hB1) |
| 12332-130I1 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 36hr | biol_rep1 (036hB1) |
| 12333-130I2 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day02 | biol_rep1 (048hB1) |
| 12334-130I3 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day03 | biol_rep1 (072hB1) |
| 12335-130I4 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day04 | biol_rep1 (096hB1) |
| 12336-130I5 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day05 | biol_rep1 (120hB1) |
| 12337-130I6 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day06 | biol_rep1 (144hB1) |
| 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 |
| 12445-132C6 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 00hr15min | biol_rep2 (015mB2) |
| 12446-132C7 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 00hr30min | biol_rep2 (030mB2) |
| 12447-132C8 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 01hr | biol_rep2 (001hB2) |
| 12448-132C9 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 02hr | biol_rep2 (002hB2) |
| 12449-132D1 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 03hr | biol_rep2 (003hB2) |
| 12450-132D2 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 06hr | biol_rep2 (006hB2) |
| 12451-132D3 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 12hr | biol_rep2 (012hB2) |
| 12452-132D4 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 18hr | biol_rep2 (018hB2) |
| 12453-132D5 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 24hr | biol_rep2 (024hB2) |
| 12454-132D6 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 36hr | biol_rep2 (036hB2) |
| 12455-132D7 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day02 | biol_rep2 (048hB2) |
| 12456-132D8 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day03 | biol_rep2 (072hB2) |
| 12457-132D9 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day04 | biol_rep2 (096hB2) |
| 12458-132E1 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day05 | biol_rep2 (120hB2) |
| 12459-132E2 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day06 | biol_rep2 (144hB2) |
| 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 |
| 12567-133H2 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 00hr15min | biol_rep3 (015mB3) |
| 12568-133H3 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 00hr30min | biol_rep3 (030mB3) |
| 12569-133H4 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 01hr | biol_rep3 (001hB3) |
| 12570-133H5 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 02hr | biol_rep3 (002hB3) |
| 12571-133H6 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 03hr | biol_rep3 (003hB3) |
| 12572-133H7 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 06hr | biol_rep3 (006hB3) |
| 12573-133H8 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 12hr | biol_rep3 (012hB3) |
| 12574-133H9 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 18hr | biol_rep3 (018hB3) |
| 12575-133I1 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 24hr | biol_rep3 (024hB3) |
| 12576-133I2 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | 36hr | biol_rep3 (036hB3) |
| 12577-133I3 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day02 | biol_rep3 (048hB3) |
| 12578-133I4 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day03 | biol_rep3 (072hB3) |
| 12579-133I5 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day04 | biol_rep3 (096hB3) |
| 12580-133I6 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day05 | biol_rep3 (120hB3) |
| 12581-133I7 | ST2 (Mesenchymal stem cells) cells, differentiation to osteocytes | day06 | biol_rep3 (144hB3) |
| 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 |
