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Osteogenic differentiation medium

Manufactured by STEMCELL
Sourced in Canada

Osteogenic differentiation medium is a cell culture medium designed to induce and support the differentiation of cells, such as mesenchymal stem cells, into osteoblasts, the cells responsible for bone formation. The medium contains specific factors and supplements that promote the development of the osteogenic lineage.

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6 protocols using osteogenic differentiation medium

1

Osteogenic and Adipogenic Differentiation

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The cells were cultured up to 80%–90% confluence and changed to the osteogenic differentiation medium or the adipogenic differentiation medium (STEMCELL Technologies, Vancouver, BC, Canada). Osteoblastic differentiation was analyzed by alizarin red staining. Adipocytic differentiation was identified by Oil Red-O staining.
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2

Murine MSCs Osteogenic Differentiation

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Murine mesenchymal stem cells (MSCs) (1.0 × 104 per well) from STEMCELL (Seattle, WA, United States) were plated in 48-well plates and cultured in MesenCultTM basal expansion medium with 10% 10x Supplement (Stemcell) for 72 h. Next, the cells were rinsed with PBS and the medium was replaced with osteogenic differentiation medium (Stemcell). MSCs cultured in expansion medium were served as the negative control. Half of the medium was changed every 3 days, and cells were harvested at 0, 5, and 7 days after induction.
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3

Isolation and Characterization of hUMSCs

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hUMSCs were extracted using the tissue block culture attachment method, as previously reported [19 (link),20 (link)]. The preparation of P5 hUMSCs was conducted in a GMP facility following good clinical practice (GCP) guidelines. The identification of hUMSCs was detected by flow cytometry (BD Biosciences, FACSAria2, USA). The antibodies used in this experiment included CD73-BV421 (cat.562430, BD Biosciences, USA), CD105-APC (cat. 562408, BD Biosciences, USA), CD90-FITC (cat.555595, BD Biosciences, USA), HLA-DR-PE (cat. 555812, BD Biosciences, USA), CD34-PerCP-Cy™5.5 (cat. 347203; BD Biosciences, USA), and CD45-FITC (cat.555482; BD Biosciences, USA). Osteogenic differentiation medium (catalog#05465, STEMCELL Technologies, China), adipogenic differentiation medium (catalog#05412, STEMCELL Technologies, China), and chondrogenic differentiation medium (catalog#05455, STEMCELL Technologies, China) were used to assess the osteogenic, adipogenic, and chondrogenic differentiation potential of hUMSCs, following the manufacturer’s instructions. Oil Red O solution (O1391, Sigma-Aldrich, USA), Alizarin Red S (A5533, Sigma-Aldrich, USA), and Alcian Blue staining solution (TMS-010-C, Sigma-Aldrich, USA) were used for adipogenic, osteogenic, and chondrogenic staining, respectively, according to the manufacturer’s instructions.
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4

Adipogenic, Osteogenic, and Chondrogenic Differentiation of ADSCs

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ADSCs cultured in DMEM/F12 medium to 90%–100% confluent were further cultured in MesenCult™ Adipogenic Differentiation Medium or Osteogenic Differentiation Medium (STEMCELL Technologies, Vancouver, Canada) for 9–10 days. The cells were fixed with 4% paraformaldehyde and stained with oil red O or alizarin red S to identify adipogenic and osteogenic differentiation, respectively.
To induce chondrogenic differentiation, ADSC pellet was incubated with MesenCult™ ACF Chondrogenic Differentiation Medium (STEMCELL Technologies, Vancouver, Canada) for 21 days. The pellets were fixed in 10% formalin and embedded with paraffin and thereafter stained with Alcian blue.
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5

Adipogenic and Osteogenic Differentiation

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Adipogenic differentiation assay was performed by culturing in the adipogenic differentiation medium (STEMCELL Technologies, Canada) for 21 days and then identified by Oil Red-O staining. Osteoblastic differentiation of CBMSCs and ASCs was induced by the osteogenic differentiation medium (STEMCELL Technologies, Canada) for 14 days and then analyzed by Alizarin Red staining.
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6

Murine MSC Osteogenic Differentiation

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Murine MSCs (1.0 × 10 4 per well) were plated in 48-well plates and cultured in MesenCult TM basal expansion medium with 10% 10x Supplement (Stemcell) for 72 h. Next, the cells were rinsed with PBS and the medium was replaced with osteogenic differentiation medium (Stemcell). MSCs cultured in expansion medium were served as the negative control. Half of the medium was changed every 3 days, and cells were harvested at 0, 5, 7 days after induction.
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