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Gibco iscove s modified dulbecco s medium imdm

Manufactured by Thermo Fisher Scientific
Sourced in United States

Gibco® Iscove's modified Dulbecco's medium (IMDM) is a cell culture medium formulated to support the growth and maintenance of a variety of mammalian cell lines, including hematopoietic cells. It is a modification of Dulbecco's Modified Eagle's Medium (DMEM) and is designed to enhance the performance of certain cell types.

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2 protocols using gibco iscove s modified dulbecco s medium imdm

1

Differentiation of BM-MSCs into Osteoblasts and Adipocytes

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BM-MSCs were induced to differentiate into osteoblasts and adipocytes using the following procedure. The induction medium for osteogenesis was Gibco® Iscove's modified Dulbecco's medium (IMDM; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS, 0.1 µM dexamethasone, 0.2 mM ascorbic acid 2-phosphate and 10 mM glycerol 2-phosphate (Sigma-Aldrich; Merck KGaA). The induction medium for adipogenesis was IMDM supplemented with 10% FBS, 1 µM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine, 10 µg/ml insulin and 60 µM indomethacin (Sigma-Aldrich; Merck KGaA). After 3 days, the culture medium was completely replaced and the medium was subsequently changed twice weekly thereafter. After the predetermined culture time had elapsed, adipocytes were stained with Oil Red O at room temperature for 30 min; the osteoblasts were identified using alkaline phosphatase (ALP) and Alizarin Red S assays were performed at room temperature in the dark for 50 min (Sigma-Aldrich; Merck KGaA).
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2

Pyrimidineamine Compound Screening Assay

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Reagents were purchased from Sigma–Aldrich (St. Louis, MO, USA) unless specified. S-Adenosyl-L-methionine (AdoMet) sulfate p-toluenesulfonate salt was purchased from Affymetrix (Santa Clara, CA, USA), Gibco Iscove’s modified Dulbecco’s medium (IMDM) was from Thermo Fisher Scientific (Waltham, MA, USA). Compound 1 was kindly provided by Genzyme (now Sanofi-Genzyme, Waltham, MA, USA). Pyrimidineamine compound stocks were made in dimethyl sulfoxide (DMSO), with an initial stock concentration of 25 or 50 mM (or the highest achievable concentration for compounds with lower solubility). Stocks were stored at –20 °C and thawed once to generate fresh solutions in DMSO for enzyme and cell-based assays. DMSO based dilution series for the enzyme assays were generated using Echo 555 acoustic liquid dispenser (Labcyte, Sunnyvale, CA, USA); final DMSO concentrations in the assays were 2%. Cell growth inhibition was determined for a 3-fold serial dilution series generated in DMSO so that final DMSO concentrations in the assay were 0.1%.
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