Mnng hos

Human osteoblasts hFOB1.19 and OS cell lines (MNNG/HOS, MG-63, S1353, U2OS, Saos-2, UMR-106) were purchased from ATCC (Rockville, USA). In RPMI-1640 complete medium containing 10% fetal bovine serum (FBS, Thermo Fisher Scientific, Shanghai, China) and 1% streptomycin (Thermo Fisher Scientific, Shanghai, China), these cells were cultured. Meanwhile, they were placed in an incubator at 37 °C with 5% CO₂ saturated humidity with the medium changed once every 2–3 days. When these cells approached fusion, they underwent 0.25% trypsinization (Thermo Fisher Scientific, Shanghai, China) and passage. It was found that RUSC1-AS1 expression was the lowest in the MG-63 cell line and the highest in the Saos-2 cell line. Therefore, MG-63 and Saos-2 became research objects in subsequent studies.

Human osteoblast cells hFOB were purchased from the American Type Culture Collection (ATCC), and NHOst were purchased from Lonza Walkersville Inc. (Walkersville, MD, USA). These cell lines were cultured in osteoblast growth medium (PromoCell) with 10% fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO, USA). Human osteosarcoma cell line KHOS was kindly provided by Efstathios Gonos (Institute of Biological Research and Biotechnology, Athens, Greece), while other cell lines U2OS, MG63, MNNG/HOS, Saos-2, and 143B were purchased from ATCC. The osteosarcoma cell lines were cultured in RPMI 1640 (GE Healthcare Life Sciences, Logan, UT, USA) supplemented with 10% FBS and 1% penicillin/streptomycin (Thermo Fisher Scientific, Waltham, MA, USA), at 37°C and 5% CO₂ in a humidified incubator. Cells were resuspended with 0.05% trypsin-EDTA before subculture.

The human osteosarcoma cell lines MNNG/HOS (CRL-1547TM, ATCC) and MG-63 (CRL-1427TM, ATCC) were from the Cell Bank of the Shanghai Institute of Biochemistry and Cell Biology, Chinese Academy of Sciences (Shanghai, China), where they were analyzed and identified. These procedures included cross-species checks, DNA identification, and quarantine. Cell lines used in this study were cultured for less than 6 months. MG-63 and MNNG/HOS were incubated in Eagle’s Minimum Essential Medium, supplemented with 10% fetal calf serum (FCS) and 1% penicillin/streptomycin mixture and both were maintained in a humidified atmosphere of 5% CO₂ and 95% air at 37°C.