Primary calvarial osteoblasts were isolated from calvaria of 5-day-old neonates using collagenase type II (50 mg/ml, Worthington, LS004176)/dispase II (100 mg/ml, Roche, 10165859001). Osteoblasts were maintained in alpha-minimal essential medium (aMEM) (Gibco) containing 10% FBS (Gibco), 2mM L-glutamine (Corning), 1% penicillin/streptomycin (Corning), and 1% nonessential amino acids (Corning) and differentiated with ascorbic acid (200 uM, Sigma, A8960). and β-glycerophosphate (10 mM, Sigma, G9422). To induce osteogenic differentiation, the growth media was supplemented with ascorbic acid (200 mM, Sigma, A8960) and beta-glycerophosphate (10 mM, Sigma, G9422).
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Isolation and Osteogenic Differentiation of Primary Calvarial Osteoblasts
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Corresponding Organization :
Other organizations : Worcester Polytechnic Institute, Baba Ghulam Shah Badshah University, University of Massachusetts Chan Medical School
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Variable analysis
independent variables
- Collagenase type II (50 mg/ml)
- Dispase II (100 mg/ml)
- Ascorbic acid (200 uM)
- Beta-glycerophosphate (10 mM)
- Osteogenic differentiation
- Alpha-minimal essential medium (aMEM) containing 10% FBS, 2mM L-glutamine, 1% penicillin/streptomycin, and 1% nonessential amino acids
- Not specified
- Not specified
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