For adipogenic differentiation, DPSCs were seeded at a density of 2×104 cells/cm2 in a 6-well plate until the cell reached 100% confluence. Then, the medium was changed to adipogenic induction medium (Cyagen Biosciences) for 3 days and subsequently maintained in adipogenic maintenance medium (Cyagen Biosciences) for 24 h. After 28 days of induction, cells were fixed with 4% PFA for 30 min and stained with Oil red O solution (Sigma-Aldrich) for 30 min to visualize lipid vacuoles.
Adipogenic maintenance medium
Manufactured by Cyagen
Sourced in China
Adipogenic maintenance medium is a cell culture medium designed to maintain the adipogenic (fat-storing) phenotype of cells during long-term culture. This medium provides the necessary nutrients and factors to support the continued differentiation and maturation of adipocytes.
Automatically generated - may contain errors
3 protocols using adipogenic maintenance medium
1
Adipogenic Differentiation of DPSCs
2
Adipogenic and Osteogenic Differentiation of hBMSCs
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The ability of hBMSCs at passage 4 to differentiate into adipocytes and osteoblasts were examined. To induce adipogenic differentiation, hBMSCs are plated in 24-well tissue culture plates with 1 ml medium per well. When the cells reached 100% confluence, they were carefully aspirated off the medium from the well, and 1 ml of adipogenic induction medium (Cyagen Biosciences, Guang Zhou, CHINA) was added. After 3 days, it was replaced with adipogenic maintenance medium (Cyagen Biosciences, Guang Zhou, CHINA), and was repeated. After 3 complete cycles of induction/maintenance, the adipocyte-induced cells were identified by staining with oil red O (Cyagen Biosciences, Guang Zhou, CHINA). To induce osteogenic differentiation, hBMSCs were plated in 24-well tissue culture plates, and when the cells reached 100% confluence, they were cultured with a osteogenic induction medium (Cyagen Biosciences, Guang Zhou, CHINA) for 21 days.The osteoblast induced cells were identified by staining with alizarin red (Cyagen Biosciences, Guang Zhou, CHINA).
3
Adipogenic Differentiation of hDPSCs
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For adipogenesis, hDPSCs reaching 80% confluence were treated with commercial adipogenic induction medium (Cyagen Biosciences, China) for 3 days, followed by adipogenic maintenance medium (Cyagen Biosciences, China) for 1 day. After completing the three cycles of induction and maintenance, the cells were incubated in adipogenic maintenance medium for another 7 days. After 21 days of induction, the adiopogenic capacity was assessed by performing oil red O (Cyagen Biosciences, China) staining. The expression of the adipogenic-related markers peroxisome proliferator-activated receptor-gamma 2 (PPARγ-2) and lipoprotein lipase (LPL) were monitored by real-time PCR and western blotting.
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