Mg 63 human osteosarcoma cell line

MG-63 human osteosarcoma cell line, human fibroblasts, and bone marrow mesenchymal stem cells were acquired from the American Type Culture Collection (ATCC), situated in Manassas, VA, USA. The MG-63 cells, human fibroblasts and bone marrow mesenchymal stem cells were grown at a temperature of 37 °C in a humidified atmosphere (5% CO₂ and 95% air environment). The MG-63 and fibroblasts culture media employed in this study consisted of Dulbecco’s Modified Eagle’s media, supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin. The bone marrow mesenchymal stem cells were supplemented with 16.5% fetal bovine serum and 1% penicillin/streptomycin. The cell lines were subjected to a minimum of four subcultures using trypsin/ethylenediaminetetraacetic acid (trypsin/EDTA) treatment and maintained in a humidified atmosphere (5% CO₂ and 95% air environment). The medium was refreshed periodically every 3 days. The cells were introduced to the 3D low attachment plates at a concentration of 5 × 10³ − 1 × 10⁴ cells/mL.

Dimethyl sulfoxide (DMSO; ≥99.7%) and 3-(4,5 dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide (MTT; 98%) were purchased from Sigma-Aldrich (St. Louis, MO, USA). The MG-63 human osteosarcoma cell line was purchased from American Type Culture Collection (ATCC, Rockville, MD, USA). Dulbecco’s Modified Eagle’s Medium, fetal bovine serum (FBS), l-glutamine, and fungizone (250 U mL⁻¹) were obtained from Pan Biotech (Aidenbach, Germany) and penicillin–streptomycin (10,000 U mL⁻¹) from Grisp (Porto, Portugal). Trypsin–ethylenediaminetetraacetic acid (EDTA) (0.25% trypsin and 1 mM EDTA) was purchased from Gibco, Life Technologies (Grand Island, NY, USA).