Project description:Ewings Sarcoma (ES) belongs to the group of bone cancers defined by the existence of a certain EWS-ETS fusion gene. In this study we use the model cell line CADO-ES1 (EWSR1-ERG fusion gene) to characterize the tumor biology of a versatile ES side-population (SP). We aim to compare SP- and non-SP-cells to identify specific characteristics of the SP which points towards a tumor driving functionality of the SP. Due to some stem cell like properties of the SP fraction a comparison to MSCs and normal fibroblasts as a control are also performed.
Project description:Murine articular chondrocytes were isolated from femoral heads of 3-week-old wild-type C57BL/6J pups as described (Gosset et al., 2008) with modifications. Following digestion, chondrocytes were harvested and cultured in complete high-glucose Dulbecco’s modified Eagle’s medium (DMEM; Gibco; #31053028) supplemented with 10% fetal bovine serum (FBS; Gibco; #10437), 2 mM L-Glutamine (Gibco; #A2916801), and 1% penicillin/streptomycin. Primary chondrocytes were seeded at a density of 50 x 104 cells/well in 12-well plates and treated with vehicle, 5 ng/ml TGFβ1 (R&D Systems; #240-B-010) and 100ng/ml BMP2 (R&D Systems; #355-BM-010) for 24 hours.
