For differentiation into adipocytes, UE7T-13 or UE7T-13/iC9 cells (5 × 103 cells) were seeded in a 96-well culture plate and cultured for 3 days at 37 °C in humidified air containing 5% CO2. Then, the medium was replaced with mesenchymal stem cell adipogenic differentiation medium (Promocell GmbH, Heidelberg, Germany) for 22 days and refreshed every 3 days. The cells that were induced to differentiate into adipocytes were stained using Oil Red O for adipocyte detection. Similarly, for differentiation into osteoblasts, UE7T-13 or UE7T-13/iC9 cells (5 × 103 cells) were seeded in a 96-well culture plate and cultured for 5 days at 37 °C in humidified air containing 5% CO2. Then, the medium was replaced with mesenchymal stem cell osteogenic differentiation medium (Promocell GmbH) for 21 days and refreshed every 3 days. The cells that were induced to differentiate into osteoblasts were stained using Alizarin Red S for osteoblast detection. These stained cells were examined under a BZ-9000 digital microscope (Keyence, Osaka, Japan).
Mesenchymal stem cell adipogenic differentiation medium
Manufactured by PromoCell
Sourced in Germany
Mesenchymal stem cell adipogenic differentiation medium is a specialized cell culture medium designed to induce and support the differentiation of mesenchymal stem cells into adipocytes, or fat cells. This medium provides the necessary growth factors and nutrients to promote the conversion of mesenchymal stem cells into mature adipose tissue cells.
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Lab products found in correlation
4 protocols using mesenchymal stem cell adipogenic differentiation medium
1
Adipocyte and Osteoblast Differentiation Assay
2
Adipogenic Differentiation of Mesenchymal Stem Cells
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For adipogenic differentiation, 1 × 105 cells per well were seeded in six-well culture plates and cultured until they reached 80–90% confluency. Subsequently, the standard culture medium was changed to the Mesenchymal Stem Cell Adipogenic Differentiation Medium (PromoCell, Heidelberg, Germany) in half of the wells. In the other half of the wells, cells were cultured in the standard culture medium to serve as controls. After 14 days of differentiation, with culture medium changed every third day, the cell monolayer was washed with PBS and fixed with Saccomanno Fixative solution for 30 min at room temperature. Subsequently, a distilled water was used to wash the cells followed with 5 min incubation with 60% isopropanol. Cell staining was performed for 3 min with Oil Red O (Sigma-Aldrich, Saint Louis, MO, USA), and then an inverted phase-contrast microscope (Olympus IX70, Olympus, Tokyo, Japan) was utilized to examine the results.
3
Osteogenic and Adipogenic Differentiation of MSCs
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rcMSCs and rbMSCs at passage 3 were used for differentiation into osteoblasts or adipocytes, as previously described7 (link),43 (link). Briefly, the cells were cultured in a mesenchymal stem cell osteogenic differentiation medium (Promocell, Heidelberg, Germany) and a mesenchymal stem cell adipogenic differentiation medium (Promocell) to induce osteogenic differentiation and adipogenic differentiation, respectively. Alizarin red S (Sigma-Aldrich, St. Louis, United States) and oil red O (Wako Pure Chemical Industries, Osaka, Japan) were used to stain the cells to confirm calcium deposition and lipid droplets, respectively.
4
Adipogenic Differentiation of Mesenchymal Stem Cells
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Adipogenic differentiation was conducted in 6-well culture plates. In total, 1 × 105 cells per well were seeded in standard culture medium and cultured until 80–90% confluent. Then, the culture medium was replaced with Mesenchymal Stem Cell Adipogenic Differentiation Medium (PromoCell, Heidelberg, Germany) in half of the wells; whereas the other half contained cells cultured as negative controls in a standard culture medium. Differentiation was conducted for 14 days and the medium was changed every 72 h. The results of the differentiation were evaluated via Oil Red O (Sigma-Aldrich, Saint Louis, MO, USA) staining. The cell monolayer was washed with PBS and fixed with Saccomanno Fixative solution for 30 min at room temperature; then, the monolayer was washed with water and incubated with 60% isopropanol for 5 min. Subsequently, the cells were stained with Oil Red O for 3 min, and the results were observed using an inverted phase-contrast microscope (Olympus IX70, Olympus, Tokyo, Japan).
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