Mouse primary BMSCs were isolated from both femurs and tibiae and cultured as described previously [43 (link)]. For osteogenic differentiation, BMSCs were cultured in osteogenic medium (α-MEM containing 10% FBS and 50 μg/mL ascorbic acid) for 7 days and then stained for ALP using a BCIP/NBT ALP color development kit (Beyotime, China). BMSCs were differentiated in osteogenic medium for 14 days for alizarin red S staining using Alizarin Red S Staining Kit (Beyotime, China) and qPCR analysis. For adipogenic differentiation, BMSCs were cultured with reagents from the MesenCult™ Adipogenic Differentiation Kit (Stemcell Technologies) for 9 days and then stained with Oil red O (Sigma) and qPCR analysis.
Mesencult adipogenic differentiation kit
Manufactured by STEMCELL
Sourced in Canada
The MesenCult Adipogenic Differentiation Kit is a cell culture medium designed to induce the differentiation of mesenchymal stem/stromal cells (MSCs) into adipocytes (fat cells) in vitro. The kit contains the necessary components to support the adipogenic differentiation process.
Automatically generated - may contain errors
Lab products found in correlation
Mesencult acf chondrogenic differentiation kit, by STEMCELL (5 mentions)
Mesencult osteogenic differentiation kit, by STEMCELL (5 mentions)
Oil red o, by Merck Group (3 mentions)
Bcip nbt alp color development kit, by Beyotime (3 mentions)
Mesencult acf chondrogenic differentiation medium, by STEMCELL (3 mentions)
Mesencult acf basal medium, by STEMCELL (3 mentions)
Mesencult osteogenic differentiation medium, by STEMCELL (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Acf enzymatic dissociation inhibition solution, by STEMCELL (2 mentions)
Alcian blue, by Merck Group (2 mentions)
Alizarin red s staining kit, by Beyotime (1 mentions)
Osteogenic differentiation kit, by Thermo Fisher Scientific (1 mentions)
Chondrogenic differentiation kit, by Thermo Fisher Scientific (1 mentions)
Mesencult chondrogenic differentiation kit, by STEMCELL (1 mentions)
Nerve growth factor (ngf), by Thermo Fisher Scientific (1 mentions)
Dbcamp, by Merck Group (1 mentions)
Neurotrophin 3, by Thermo Fisher Scientific (1 mentions)
Brain derived neurotrophic factor (bdnf), by R&D Systems (1 mentions)
Fibroblast growth factor 2 (fgf2), by Merck Group (1 mentions)
L alanyl l glutamine, by Thermo Fisher Scientific (1 mentions)
21 protocols using mesencult adipogenic differentiation kit
1
Isolation and Differentiation of Mouse BMSCs
2
Multilineage Differentiation of Mesenchymal Stem Cells
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The differentiation capacities of MSCs into adipogenic, osteogenic, or chondrogenic lineages were performed using following kits: the MesenCult Adipogenic Differentiation kit (Stemcell, Canada), Osteogenic Differentiation kit (Gibco), or Chondrogenic Differentiation kit (Gibco) according to the manufacturer’s procedures as we previously reported [9 (link), 11 (link)]. In brief, MSCs were seeded into 12-well plates at a density of 5 × 104 cells per well and cultured until they were approximately 90–100% confluent. Then, the medium was replaced by corresponding differentiation medium and refreshed every 4 days. Oil red O staining, alizarin red S staining, and Alcian blue staining were performed to measure adipogenesis, osteogenesis, and chondrogenesis after cultured for 2 weeks, respectively. Finally, the stained cells were observed under a microscope. In addition, total RNA was extracted when the corresponding differentiation assays were completed.
3
Tri-lineage Differentiation of Cryopreserved Cells
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Tri-lineage differentiation was assessed for cells seeded directly from cryopreservation. Adipogenic differentiation was performed using the MesenCult Adipogenic Differentiation Kit (Stemcell Technologies) according to the manufacturer’s instructions. Similarly, chondrogenic differentiation was done using the MesenCult Chondrogenic Differentiation Kit (Stemcell Technologies) according to the manufacturer’s instructions. The osteogenic differentiation protocol consisted of 25–28 day culture on hMSC medium with the addition of β-glycerolphosphate (0.01 M) and dexamethasone (10−8M). For adipogenic differentiation, presence of lipid vacuoles was demonstrated using Oil Red O staining. For chondrogenic differentiation, presence of aggrecan was demonstrated using Alcian Blue staining. For osteogenic differentiation, calcium deposits were stained with Alizarin Red.
4
Differentiation of Neural Crest Cells
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For peripheral neuron differentiation, NCCs were cultured in neuron differentiation media containing DMEM/F12, N2 supplement (Gibco), BDNF (10 ng/mL, R&D Systems), GDNF (10 ng/mL, R&D Systems), NGF (10 ng/mL, Peprotech), neurotrophin-3 (10 ng/mL, Peprotech), sodium l -ascorbic acid salt (200 μM) and dbcAMP (0.5 mM, Sigma) for 12–14 days. The medium was changed every 2 days. For mesenchymal differentiation, hiPSC-NCCs were cultured in media containing DMEM/F12, 10% FBS (Gibco), 1% penicillin-streptomycin (Corning), 1% l -alanyl-l -glutamine (Gibco), and 2-mercaptoethanol (0.1 mM, Sigma). The differentiated cells were passaged every 4 to 5 days and the media were changed every 2 days. Adipoblast, osteoblast, and chondroblast differentiation were performed according to the manufacturer’s directions using MesenCult Osteogenic Differentiation Kit (catalog 05465), MesenCult Adipogenic Differentiation Kit (catalog 05412), and MesenCult-ACF Chondrogenic Differentiation Kit (catalog 05455) (STEMCELL Technologies), respectively. For corneal keratocyte differentiation, NCCs were cultured in matrigel-coated plates and keratocyte differentiation media containing DMEM/F12, FGF2 (10 ng/mL), ascorbic acid-2-phosphate (1 mM, Sigma), 1% ITS, and 1% NEAA.
5
Adipogenic and Osteogenic Differentiation of SCAP-S and DPSCs
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SCAP-Ss and DPSCs at indicated passages were seeded at a density of 5 × 104/cm2 in the aforementioned culture medium as reported [2 (link), 8 (link)]. When cells reached 80% confluency, the medium was changed into adipogenic differentiation medium (MesenCult Adipogenic Differentiation Kit, Stem Cell Technologies) or osteogenic differentiation medium (MesenCult Osteogenic Differentiation Kit, Stem Cell Technologies). The differentiation medium was changed every 3.5 days as we described previously [2 (link), 3 (link), 23 (link)]. 21 days later, the SCAP-S and DPSC-derived cells were stained with Oil Red O or Alizarin Red staining buffer, respectively. Then, the cells were photographed with a Nikon Eclipse Ti-U microscope (Nikon, Tokyo, Japan).
6
Osteogenic and Adipogenic Differentiation of MSCs
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MSCs were orientally induced under passage 8 when they reached 80% confluence. For osteogenic induction, the cells were washed with 1 × dPBS and then cultured with OsteoMAX-XF Differentiation Medium (catalog number SCM121, EMD Millipore, MD, USA) supplemented with 1% penicillin–streptomycin. The osteogenic medium was half-changed on day 2, 4, 7, 9, and 11, according to the previous study [25 (link)]. MesenCult Adipogenic Differentiation Kit (catalog number 05412, STEMCELL Technologies, Vancouver, Canada) supplemented with 1% penicillin–streptomycin was used for adipogenic induction, and all the culture medium was replaced at the same time point mentioned above. The samples were collected on day 0, 7, and 14 for the experiments.
7
Multilineage Differentiation of MSCs
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Example 13
Adipocytes, osteocytes, and chondrocytes were generated from MSCs. For adipogenic lineage differentiation, MSCs between passages 3-4 were plated on 6-well plates with MesenCult-ACF Basal Medium (Stem Cell Technologies, Cat. #05449) at 4-5×105 cells per well. Differentiation was performed using the MesenCult Adipogenic Differentiation Kit (Stem Cell Technologies, Cat. #05412) according to manufacturer's protocol. Media changes were conducted every 3-4 days until day 13. For osteogenic lineage differentiation, MSCs between passages 3-4 were plated on a 6-well plate with MesenCult-ACF Basal Medium (Stem Cell Technologies) at 3-4×104 cells per well. Differentiation was performed using the MesenCult Osteogenic Differentiation medium (Stem Cell Technologies, Cat. #05465) according to manufacturer's protocol. Medium changes were conducted every 3-4 days until day 13. For 3D pellet chondrogenic differentiation, MSCs were first released from T25 flasks using ACF Enzymatic Dissociation/Inhibition Solutions (Stem Cell Technologies, Cat. #05426) and collected in polypropylene tubes at 2.5-3×106 cells per tube with MesenCult-ACF Chondrogenic Differentiation Medium (Stem Cell Technologies, Cat. #05455) according to manufacturer's protocol. Medium changes were conducted every 3-4 days until day 13.
8
Isolation and Differentiation of Mouse Primary BMSCs
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Mouse primary BMSCs were isolated and cultured as described previously.70 (link) To evaluate BMSC proliferation, the number of attached cells was assayed by the Cell Counting Kit-8 (Beyotime) assay according to the manufacturer’s instructions. The optical density at 450 nm was determined with a microplate reader (PerkinElmer). For osteogenic differentiation, BMSCs were cultured in osteogenic medium (α-MEM containing 10% FBS and 50 μg·mL−1 ascorbic acid) for 7 days and then stained for ALP using a BCIP/NBT Alp color development kit (Beyotime, China) or cultured in osteogenic medium for 7 days followed by mineralization-inducing medium (osteogenic medium plus 2.5 mmol·L−1 β-glycerophosphate for 7 days and then subjected to alizarin red S (40 mmol·L−1, pH 4.2) (Sigma) staining. For adipogenic differentiation, BMSCs were cultured with reagents from the MesenCult™ Adipogenic Differentiation Kit (STEMCELL Technologies) for 9 days and then stained with Oil Red O (Sigma).
9
Multilineage Differentiation of Mesenchymal Stem Cells
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Adipocytes, osteoblasts, and chondrocytes were generated from MSCs. For adipocyte differentiation, MSCs between passages 3 – 4 were plated on 6-well plates with MesenCult-ACF Basal Medium (StemCell Technologies, Cat. # 05449) at 4 – 5 3 105 cells per well. Differentiation was performed using the MesenCult Adipogenic Differentiation Kit (StemCell Technologies, Cat. # 05412) according to the manufacturer’s protocol. Media changes were done every 3 – 4 d until 13 d. For osteogenic lineage differentiation, MSCs between passages 3 – 4 were plated on a 6-well plate with MesenCult-ACF Basal Medium (StemCell Technologies) at 3 – 4 × 104 cells per well. Differentiation was performed using MesenCult Osteogenic Differentiation medium (StemCell Technologies, Cat. # 05465) according to the manufacturer’s protocol. Medium changes were done every 3 – 4 d until 13 d. For 3-D pellet chondrogenic differentiation, MSCs were first released from T25 flasks using ACF Enzymatic Dissociation/Inhibition Solution (StemCell Technologies, Cat. # 05426) and collected in polypropylene tubes at 2.5 – 3 × 106 cells per tube with MesenCult-ACF Chondrogenic Differentiation Medium (StemCell Technologies, Cat. # 05455) according to the manufacturer’s protocol. Medium changes were done every 3 – 4 days until 13 d.
10
Murine BMSC Osteogenic and Adipogenic Differentiation
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Primary BMSCs were isolated from both femurs and tibiae of 6-month-old mice and cultured as described previously [28 ]. For osteogenic differentiation, BMSCs were cultured in osteogenic medium (α-MEM containing 10% FBS and 50 μg/mL ascorbic acid) for 7 days and then stained for ALP using a BCIP/NBT ALP color development kit (Beyotime, China). For adipogenic differentiation, BMSCs were cultured with reagents from the MesenCult™ Adipogenic Differentiation Kit (Stemcell Technologies) for 9 days and then stained with Oil Red O (Sigma).
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