The MSCs were seeded into 6-well plates in a density of 20,000 cells/cm2. While cells were grown to confluence, they were transferred to adipogenic induction medium containing LG-DMED and adipogenic stimulatory supplement (Stem Cell Technologies, Hangzhou, China) and the system was cultured for 21 days. The medium was changed every 3 days. R-2HG at a concentration of 0–1.5 mM was added to the adipogenic induction medium. The adipogenic differentiation was mesured by cellular accumulation of large lipid vacuoles that are stained with oil red O (Sigma-Aldrich).
Adipogenic stimulatory supplement
Manufactured by STEMCELL
Sourced in China
The Adipogenic stimulatory supplement is a laboratory reagent designed to promote the differentiation of cells into adipocytes (fat cells) in vitro. It provides the necessary factors to stimulate and support the adipogenic differentiation process.
Automatically generated - may contain errors
Lab products found in correlation
Lg dmed, by STEMCELL (4 mentions)
Oil red o, by Merck Group (4 mentions)
Mesencult msc basal medium, by STEMCELL (3 mentions)
Ascorbic acid, by STEMCELL (2 mentions)
Osteogenic stimulatory supplement, by STEMCELL (1 mentions)
Dexamethasone (dex), by STEMCELL (1 mentions)
β glycerophosphate, by STEMCELL (1 mentions)
Stempro chondrocyte differentiation basal medium, by Thermo Fisher Scientific (1 mentions)
Stempro chondrogenesis supplement, by Thermo Fisher Scientific (1 mentions)
Adipogenic induction medium, by STEMCELL (1 mentions)
Pen strept, by Thermo Fisher Scientific (1 mentions)
Pericyte medium, by ScienCell (1 mentions)
Rabbit osterix antibody, by Abcam (1 mentions)
Rabbit perilipin antibody, by Merck Group (1 mentions)
Tgf β3, by Thermo Fisher Scientific (1 mentions)
Ascorbic acid, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Dexamethasone (dex), by Merck Group (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
Eclipse ti, by Nikon (1 mentions)
9 protocols using adipogenic stimulatory supplement
1
Adipogenic Differentiation of MSCs with R-2HG
2
Differentiation of human BM-derived MSCs
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To qualify isolated cells from human BM as hMSCs, cells were induced to differentiation into adipogenic, osteogenic and chondrogenic lineages. Briefly, for adipogenic differentiation, 5x103 hMSCs/cm2 were exposed to Complete MesenCult Adipogenic Medium containing MesenCult MSC Basal Medium (Stemcell) and 10% Adipogenic Stimulatory Supplement (Stemcell) for 3 weeks. For osteogenic differentiation, 2x105 cells/cm2 incubated with Complete MesenCult Osteogenic Medium including MesenCult MSC Basal Medium, Osteogenic Stimulatory Supplement, β-Glycerophosphate, Dexamethasone, Ascorbic acid (all from Stemcell) for 5 weeks. For chondrogenic differentiation, 7.5x106 cells/cm2 were cultured for 3 weeks with Stempro Chondrocyte Differentiation Basal Medium (Gibco) containing 10% Stempro Chondrogenesis Supplement (Gibco). Standard histochemical staining methods were applied. Osteogenic, adipogenic and chondrogenic differentiation were confirmed by Toluidine Blue, Oil Red O, Alcian Blue staining, respectively.
3
Adipogenic Differentiation of 3T3-L1 Cells
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MCSs were seeded at p2 at 2 × 104 cells/cm2 and at 80% confluence the Complete Medium was replaced with Adipogenic Induction Medium (MesenCult Basal Medium supplemented with 20% AdipogenicStimulatory Supplement (StemCell Technologies) and 1% Pen-Strept (Life Technologies) we followed the manufacturer’s recommendations.
For adipogenic differentiation of 3T3-L1 cells, two days after confluence the medium was changed to differentiation medium I containing 1 μM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine, and 1 μg/ml insulin in DMEM containing 10% of FBS. Two days after the induction of differentiation the medium was changed do differentiation medium II containing only 1 μg/ml insulin in the presence of 10% of FBS. After 4 days of differentiation the medium was changed to differentiation medium III containing only 10% of FBS.
For adipogenic differentiation of 3T3-L1 cells, two days after confluence the medium was changed to differentiation medium I containing 1 μM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine, and 1 μg/ml insulin in DMEM containing 10% of FBS. Two days after the induction of differentiation the medium was changed do differentiation medium II containing only 1 μg/ml insulin in the presence of 10% of FBS. After 4 days of differentiation the medium was changed to differentiation medium III containing only 10% of FBS.
4
Culturing Primary PDGFRβ+ Cells and Satellite Cells
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The primary PDGFRβ+ cells were grown in pericyte medium (ScienCell, 1201), myogenic differentiation medium (DMEM with 2% horse serum and 1% penicillin/streptomycin) or adipogenic differentiation medium (MesenCult Basal Medium supplemented with Adipogenic Stimulatory Supplement, STEMCELL, 05501 and 05503) at 37 °C with 5% CO2.
The satellite cells were grown in satellite cell medium (DMEM with 10% FBS, 10% horse serum and 1% penicillin/streptomycin) at 37 °C with 5% CO2 for 16 h.
The satellite cells were grown in satellite cell medium (DMEM with 10% FBS, 10% horse serum and 1% penicillin/streptomycin) at 37 °C with 5% CO2 for 16 h.
5
Adipogenic Differentiation of hMSCs
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For adipogenic differentiation, the hMSC line was harvested at passage 3 (p3) and seeded into 24-well plates at a density of 5 × 103 cells/cm2. Cells were treated with complete MesenCult adipogenic medium containing MesenCult MSC basal medium (Stemcell) and 10% adipogenic stimulatory supplement (Stemcell) for 21 days; adipogenic differentiation was confirmed by Oil Red O staining.
6
Isolation and Differentiation of Bone Marrow Stromal Cells
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Long bones were collected and the BM was flushed by centrifugation to obtain marrow stromal cells.35 Following red blood cell (RBC) lysis, 105 to 106 cells were plated in MesenCult MSC Basal Medium supplemented with MesenCult Mesenchymal Stem Cell Stimulatory Supplement (STEMCELL Technologies), glutamine and antibiotics. Adherent cells were analyzed after 24 hours and colonies were visualized after 2 weeks. To promote differentiation, cells were cultured to confluence and then the media was supplemented with either 50 μg/mL ascorbic acid (Sigma), 7 mM β‐glycerophosphate (Sigma), 10−8 dexamethasone (Sigma) for osteoblastogenesis, 20 ng/mL TGF‐β3 (PeproTech), 200 mM ascorbic acid for chondrogenesis or Adipogenic Stimulatory Supplement (Stem Cell Technologies) for adipogenesis. Differentiation media was changed twice a week and cells were cultured up to 4 weeks. Immunofluorescence was performed using rabbit perilipin antibody (Sigma, 5 μg/mL), goat collagen II antibody (Santa Cruz, 1:250) or rabbit osterix antibody (Abcam, 1:600) and Alexa Fluor 350, 633 or 647 secondary antibodies (Molecular Probes, 1:400) and imaged using an inverted Eclipse Ti (Nikon) or LSM 700 laser scanning confocal (Zeiss) microscope. Nuclei were stained using 4′,6‐diamidino‐2‐phenylindole (DAPI) (Sigma, 1:1000).
7
Adipogenic Differentiation of MSCs
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The MSCs were seeded into 6-well plates in a density of 20,000 cells/cm 2 . While cells were grown to con uence, they were transferred to adipogenic induction medium containing LG-DMED and adipogenic stimulatory supplement (Stem Cell Technologies, Hangzhou, China) and the system was cultured for 21 days. The medium was changed every 3 days. R-2HG at a concentration of 0-1.5 mM was added to the adipogenic induction medium. The adipogenic differentiation was mesured by cellular accumulation of large lipid vacuoles that are stained with oil red O (Sigma-Aldrich).
8
Adipogenic Differentiation of MSCs
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The MSCs were seeded into 6-well plates in a density of 20,000 cells/cm 2 . While cells were grown to con uence, they were transferred to adipogenic induction medium containing LG-DMED and adipogenic stimulatory supplement (Stem Cell Technologies, Hangzhou, China) and the system was cultured for 21 days. The medium was changed every 3 days. R-2HG at a concentration of 0-1.5 mM was added to the adipogenic induction medium. The adipogenic differentiation was mesured by cellular accumulation of large lipid vacuoles that are stained with oil red O (Sigma-Aldrich).
9
Adipogenic Differentiation of MSCs
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The MSCs were seeded into 6-well plates in a density of 20,000 cells/cm 2 . While cells were grown to con uence, they were transferred to adipogenic induction medium containing LG-DMED and adipogenic stimulatory supplement (Stem Cell Technologies, Hangzhou, China) and the system was cultured for 21 days. The medium was changed every 3 days. R-2HG at a concentration of 0-1.5 mM was added to the adipogenic induction medium. The adipogenic differentiation was mesured by cellular accumulation of large lipid vacuoles that are stained with oil red O (Sigma-Aldrich).
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