Cells were grown in osteogenic induction medium (Gibco) for 21 days. Calcium deposition was shown by alizarin red staining (Sigma, Steinheim, Germany).
Osteogenic induction medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
Osteogenic induction medium is a specialized cell culture medium designed to promote the differentiation of cells into osteoblasts, which are responsible for bone formation. The medium contains a proprietary blend of nutrients, growth factors, and other components that support the osteogenic differentiation process. This product is intended for use in a controlled laboratory setting by trained professionals.
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Lab products found in correlation
Alizarin red s, by Merck Group (2 mentions)
Alizarin red, by Merck Group (1 mentions)
Chondrogenic medium, by Thermo Fisher Scientific (1 mentions)
Adipogenic medium, by Thermo Fisher Scientific (1 mentions)
Flow cytometry analyzer, by Beckman Coulter (1 mentions)
Oil red o, by Merck Group (1 mentions)
Adipogenic induction medium, by Thermo Fisher Scientific (1 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (1 mentions)
Sc 16143, by Santa Cruz Biotechnology (1 mentions)
Sc 292097, by Santa Cruz Biotechnology (1 mentions)
8 protocols using osteogenic induction medium
1
Osteogenic Differentiation Monitoring
2
Multilineage Differentiation and Characterization of Umbilical Stem Cells
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After the isolation of USCs, we used the Alizarin Red Staining, Oil Red O staining, and Alcian blue staining to verify their multilineage differentiation potential. For osteogenic differentiation, USCs were incubated with an osteogenic induction medium (Gibco). On day 21, after the induction, the cells were stained with Alizarin Red S (Sigma, USA) for observation. For adipogenic differentiation, USCs were incubated with an adipogenic medium (Gibco). On day 21, after the induction, USCs were stained with Oil Red O (Sigma) for observation. For the chondrogenic differentiation, USCs were incubated with a chondrogenic medium (Gibco). After 21 days of induction, allicin blue staining (Sigma) was performed for observation. We also performed the flow cytometry analysis to detect the stem-cell surface markers (CD44, CD73, CD90, CD105, CD31, CD34, and CD45). The USCs were incubated with these antibodies (BD, USA), washed with PBS, and analyzed with a flow cytometry analyzer (Beckman, USA).
3
Osteogenic Differentiation of Cells
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The cells were grown to 80% confluence in a 6-well plate and incubated with osteogenic induction medium (Gibco, USA), which was refreshed every 3 days. After 28 days of induction, the cells were stained with Alizarin Red S (Sigma, USA) to detect calcified extracellular matrix.
4
Adipogenic and Osteogenic Differentiation of ADSCs
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After the cell confluence of ADSCs at passage 3 reached 80%. For osteogenic induction, ADSCs were cultured in osteogenic induction medium (Gibco) for 21 days, and the cultured cells were subsequently stained with Alizarin Red S to detect the deposition of calcified matrix. ADSCs were cultured in adipogenic differentiation medium to identify the adipogenic differentiation ability. After 14 days, oil red O staining [27 (link)] was carried out to characterize the morphological changes under an inverted phase contrast microscope. The expression of ADSC surface marker antigens CD49d, CD90, CD105, CD34, CD45, and CD106 were identified by flow cytometry.
5
Osteogenic Differentiation of USCs
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Cultured USCs at passage 4 were grown to 80% confluence and subsequently incubated with osteogenic induction medium (Gibco, USA). After 21 days of induction, the cells were stained with Alizarin Red S to detect the deposition of calcified matrix.
6
Multi-lineage Differentiation of UC-MSCs
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The multi-lineage differentiation potential of human UC-MSCs was checked by adipogenic, osteogenic and neural-like differentiation assays at the fourth passage. Adipogenesis was induced by adipogenic induction medium (Gibco) for 14 days and confirmed by Oil red O staining to show intracellular lipid accumulation. Osteogenesis was induced by osteogenic induction medium (Gibco) for 28 days and calcium deposition was shown by Alizarin red staining. For neural-like differentiation, UC-MSCs seeded on poly-L-lysine-coated coverslips in a 24-well culture plate were treated with pre-induction medium containing 10-7 M all-trans-retinoic acid (ATRA; Sigma-Aldrich, St. Louis, MO, USA) and 10 ng/ml bFGF (Gibco) for 24 h and then with modified MNM medium for 36 h. The cells were co-incubated with the anti-NSE antibody (1:100, sc-292097, Santa Cruz Biotechnology) and the anti-NF-M antibody (1:100, sc-16143, Santa Cruz Biotechnology) to confirm their differentiation into neural-like cells.
7
Osteogenic Differentiation of MSCs
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For osteogenic differentiation 50,000 MSCs per well were seeded in 6 well plates in triplicate in expansion medium. After 24 hours (at 90% confluency) osteogenic differentiation was promoted by treating MSC cultures with osteogenic induction medium (ThermoScientific, USA) for 3 weeks while MSCs maintained in expansion medium for 3 weeks were used as controls.
8
Osteogenic Induction of Stem Cells
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After culturing for 7 days, 1 mL osteogenic induction medium (ThermoFisher Scientific Inc., USA) was added into the plate instead of proliferation culture medium. The compositions of medium were 10 μL penicillin-streptomycin solution, 10 μL glutamine, 2 μL ascorbic acid, 10 μL glycerophosphate, 0.1 μL dexamethasone, 100 μL fetal calf serum (FBS) and 875 μL basal medium. The media was changed every 3 days, cell morphology was observed by optical microscope at the same time.
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