Osteogenic differentiation was conducted as we previously described with several modification (10 (link)). In details, ND-ASCs and T2DM-ASCs were seeded at a density of 2×104 cells/cm2 at 37℃, 5% CO2. When cells reached 80% confluent, the supernatant was discarded and changed into osteogenic differentiation medium containing 100 nM dexamethasone, 50 μM L-ascorbic acid 2-phosphate, and 10 mM β-glycerophosphate in Serum-free human MSC medium (Yocon). The medium was subsequently changed twice a week. After 28 days, osteogenic differentiation was assessed by von Kossa staining (Abcam). The phase contrast image was photographed with Nikon ElipseTi-U microscope (Nikon, Tokyo, Japan).
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Osteogenic Differentiation of ASCs
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Ascorbic acid 2 phosphate
Ascs
Cells
Dexamethasone
Human
Microscope
Osteogenic
Phase contrast
Serum
Yocon
β glycerophosphate
Corresponding Organization :
Other organizations : Tianjin Medical University, Tianjin First Center Hospital, Nankai University, Shanghai Clinical Research Center
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Variable analysis
independent variables
- Cell type: ND-ASCs (non-diabetic adipose-derived stem cells) and T2DM-ASCs (type 2 diabetes mellitus adipose-derived stem cells)
- Osteogenic differentiation, assessed by von Kossa staining
- Cell seeding density: 2 × 10^4 cells/cm^2
- Culture conditions: 37°C, 5% CO2
- Osteogenic differentiation medium: 100 nM dexamethasone, 50 μM L-ascorbic acid 2-phosphate, and 10 mM β-glycerophosphate in Serum-free human MSC medium (Yocon)
- Medium change frequency: Twice a week
- Culture duration: 28 days
- No positive or negative controls were explicitly mentioned in the provided information.
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