Synovial MSCs were transferred to a 15 ml tube (Corning Falcon, NY) and cultured in chondrogenic induction medium containing 10 ng/ml transforming growth factor‐β3 (Miltenyi Biotec, Tokyo, Japan), 500 ng/ml bone morphogenetic protein 2 (Medtronic, TN), 10−7 M dexamethasone (Sigma–Aldrich), 50 µg/ml ascorbate‐2‐phosphate, 40 µg/ml proline, 100 µg/ml pyruvate, and 50 mg/ml ITS Premix (Corning); this induction medium was changed every 3–4 days. After 21 days, images of cartilage pellets were obtained with a Leica M165 FC stereoscopic microscope (Leica, Wetzlar, Germany), and the pellets were weighed. The optimal cell number was determined by culturing an initial 250k, 125k, 63k, or 31k synovial MSCs; 125k cells were used for further analyses.
Bone morphogenetic protein 2
Manufactured by Medtronic
Sourced in Japan
Bone morphogenetic protein 2 is a laboratory reagent used in research and development. It is a signaling protein that plays a role in bone and cartilage formation.
Automatically generated - may contain errors
Lab products found in correlation
Transforming growth factor β3, by Miltenyi Biotec (3 mentions)
Dexamethasone (dex), by Merck Group (2 mentions)
15 ml tube, by BD (2 mentions)
Its premix, by Corning (1 mentions)
15 ml tube, by Corning (1 mentions)
Indomethacin, by Fujifilm (1 mentions)
Dexamethasone (dex), by Fujifilm (1 mentions)
Alizarin red staining, by Merck Group (1 mentions)
Isobutylmethylxanthine, by Merck Group (1 mentions)
Oil red o staining, by Muto Pure Chemicals (1 mentions)
Ascorbic acid 2 phosphate, by Fujifilm (1 mentions)
β glycerophosphate, by Merck Group (1 mentions)
Safranin o, by Fujifilm (1 mentions)
3 protocols using bone morphogenetic protein 2
1
Chondrogenic Induction of Synovial MSCs
2
Multilineage Differentiation Potential of Synovial MSCs
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For adipogenesis, synovial MSCs were plated at 100 cells/60-cm2 dish and cultured for 14 days in culture medium. The adherent cells were cultured in adipogenic induction medium supplemented with 100 nM dexamethasone, 0.5 mM isobutyl-methylxanthine (Sigma-Aldrich), and 50 mM indomethacin (Wako) for an additional 21 days. Adipocytes were stained with oil red-o staining (Muto Pure Chemicals, Tokyo, Japan).
For calcification, 100 cells were transferred to a 60-cm2 dish and cultured for 14 days in culture medium. The adherent cells were cultured in calcification induction medium containing 50 μg/mL ascorbic acid 2-phosphate (Wako), 10 nM dexamethasone (Wako), and 10 mM β-glycerophosphate (Sigma-Aldrich). After 21 days, calcification was assessed by alizarin red staining (Merck Millipore, Billerica, MA, USA).
For chondrogenesis, 250,000 synovial MSCs were transferred to a 15-mL tube (BD Falcon) and cultured in chondrogenic induction medium containing 10 ng/mL transforming growth factor-β3 (Miltenyi Biotec K.K., Tokyo, Japan) and 1 μg/mL bone morphogenetic protein 2 (Medtronic, TN, USA), which was changed every 3–4 days. After 21 days, cartilage pellets were weighed and evaluated histologically.
For calcification, 100 cells were transferred to a 60-cm2 dish and cultured for 14 days in culture medium. The adherent cells were cultured in calcification induction medium containing 50 μg/mL ascorbic acid 2-phosphate (Wako), 10 nM dexamethasone (Wako), and 10 mM β-glycerophosphate (Sigma-Aldrich). After 21 days, calcification was assessed by alizarin red staining (Merck Millipore, Billerica, MA, USA).
For chondrogenesis, 250,000 synovial MSCs were transferred to a 15-mL tube (BD Falcon) and cultured in chondrogenic induction medium containing 10 ng/mL transforming growth factor-β3 (Miltenyi Biotec K.K., Tokyo, Japan) and 1 μg/mL bone morphogenetic protein 2 (Medtronic, TN, USA), which was changed every 3–4 days. After 21 days, cartilage pellets were weighed and evaluated histologically.
3
Chondrogenic Differentiation of MSCs with and without Trisomy 7
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MSCs from six donors with and without trisomy 7 were harvested at P2 in a cell‐dissociation buffer. A total of 2.5 × 105 cells were transferred to a 15‐mL tube (BD Falcon, New Jersey) and cultured in chondrogenic induction medium containing 10 ng/mL transforming growth factor‐β3 (Miltenyi Biotec GmbH, Bergisch Gladbach, Germany) and 1 μg/mL bone morphogenetic protein 2 (Medtronic, Memphis, Tennessee); the medium was changed every 3 to 4 days. After 21 days, chondrogenic differentiated cells were analyzed by staining with safranin O (Fujifilm Wako), and DNA was extracted.22 Chondrogenic ability was evaluated by wet weight, as described previously.23 , 24
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