Human osteosarcoma cell lines (MG-63 and SAOS-2) were obtained from the Korean Cell Line Bank (Seoul, Korea). MG-63 and SAOS-2 cells were cultured in RPMI 1640 (Lonza, Basel, Switzerland) with 10% FBS maintained at 37 °C in a humidified atmosphere containing 5% carbon dioxide. The human fetal osteoblastic hFOB 1.19 cell line was cultured according to established ATCC protocols. Mycoplasma infection was tested regularly using a PCR-based kit (MP0040; Sigma, St. Louis, MO, USA). siRNA duplexes directed against human Axin2 (sc-35087) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Niclosamide (2′,5-dichloro-4′-nitrosalicylanilide) was purchased from Cayman (Ann Arbor, MI, USA), and pyrvinium (6-(dimethylamino)-2-[2-(2,5-dimethyl-1-phenyl-1H-pyrrol-3-yl) ethenyl]-1-methyl-4,4′-methylenebis[3-hydroxy-2-naphthalenecarboxylate] (2:1)-quinolinium) was purchased from Sigma-Aldrich (Burlington, MA, USA). Niclosamide (Cayman, Ann Arbor, MI, USA) and pyrvinium pamoate (Sigma, St. Louis, MO, USA) were solubilized in DMSO for in vitro experiments.
Mg 63
Manufactured by Lonza
Sourced in Switzerland
The MG-63 is a cell line derived from human osteosarcoma. It is commonly used in various cell culture applications for research purposes.
Automatically generated - may contain errors
Lab products found in correlation
Saos2, by Lonza (2 mentions)
Rpmi 1640, by Lonza (2 mentions)
Niclosamide, by Cayman Chemical (1 mentions)
Pyrvinium pamoate, by Merck Group (1 mentions)
Sirna duplexes, by Santa Cruz Biotechnology (1 mentions)
Mp0040, by Merck Group (1 mentions)
Saos2, by Korean Cell Line Bank (1 mentions)
Hfob1, by American Type Culture Collection (1 mentions)
Sterile distilled water, by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Lonza (1 mentions)
Dulbecco modified eagle medium (dmem), by Lonza (1 mentions)
Rpmi 1640 medium, by Lonza (1 mentions)
Saos 2, by American Type Culture Collection (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
Sjsa 1, by American Type Culture Collection (1 mentions)
Crl1554, by American Type Culture Collection (1 mentions)
Crl 8303, by American Type Culture Collection (1 mentions)
Crl 1427, by American Type Culture Collection (1 mentions)
4 protocols using mg 63
1
Osteosarcoma Cell Lines and Reagents
2
Establishment of MTX-Resistant Osteosarcoma Cell Lines
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The human osteoblast hFOB1.19 (catalog: CRL-11372), human osteosarcoma cell lines U2OS (catalog: HTB-96), and MG63 (catalog: CRL-1427) were sourced from the American Type Culture Collection (Manassas, VA, USA). U2OS and MG63 were cultured in Dulbecco’s modified Eagle medium (catalog: BE12-604F, Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (catalog: F7524, Sigma-Aldrich, St. Louis, MO, USA). hFOB1.19 cells were cultivated in hFOB1.19 complete medium (catalog: EP-ML-0353, Elabscience, Wuhan, China). All cells were kept at 37 °C in a 5% CO2 environment.
Parental U2OS and MG63 cells were initially exposed to 3 ng/mL (0.01 µM) MTX [15 (link)]. Subjecting the parental cell lines to stepwise raised MTX concentrations, MTX-resistant subclones were produced.
Parental U2OS and MG63 cells were initially exposed to 3 ng/mL (0.01 µM) MTX [15 (link)]. Subjecting the parental cell lines to stepwise raised MTX concentrations, MTX-resistant subclones were produced.
3
Evaluating Nanoparticle Cytotoxicity in Osteosarcoma and Mesenchymal Stem Cells
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MG-63, an osteosarcoma cell line (Lonza, UK) and human bone marrow derived primary mesenchymal stem cells (less than 5 passages) (Lonza, UK) were seeded in expansion media consisting of 4.5 gL−1 glucose Dulbecco’s Modified Eagle’s medium (Lonza, UK) supplemented with 10% foetal bovine serum, 1% Penicillin/Streptomycin (antibiotics and antimycotics) and 1% L-glutamine in well plates at ~80% confluency and allowed to attach overnight before addition of nanoparticle suspensions. hMSCs were adherence selected and tested for stemness using standard trilineage differentiation and flow cytometry for CD markers. Aqueous suspensions of the nanoparticles were mixed with cell culture media at 10% v/v and then added to cells. For the control samples, equivalent amount of sterile distilled water (negative control) or sterile cobalt chloride solution (positive control; Sigma- Aldrich, UK) at a concentration of 500 μM were mixed with the cell culture media and added to cells. The cation/MNP concentrations were maintained constant between comparisons and cells were incubated with the additives for 72 h before being assessed.
4
Murine and Human Osteosarcoma Cell Lines
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The murine osteosarcoma MOS-J cell line, called MOS-J/Native in this study, derived from a spontaneous C57BL/6 mouse osteosarcoma, was provided by Prof. L. Shultz [21 ]. Two subclones derived from this cell line were also used in the experiments [6 (link)]. The first clone, MOS-J/PG1, revealed a high proliferation rate in vitro in contrast to the second, MOS-J/A3N. These clones, as well as the MOS-J/Native, were grown in RPMI1640 medium (Lonza, Walkersville, MD, USA) supplemented with 5% fetal bovine serum (FBS; Hyclone, Logan, UT, USA) and a mix of 100 U/mL of penicillin and 100 µg/mL of streptomycin (Lonza). The human osteosarcoma cell lines G292 (clone A141B1), KHOS/NP (R-970-5) named HOS in the manuscript, 143B, MG63, SJSA-1, and SaOS2 were purchased at the American Type Culture Collection (ATCC, Manassas, VA, USA), respectively, under references CRL-1423, CRL-1554, CRL-8303, CRL-1427, CRL-2098, and HTB-85. The CAL-72 cell line was purchased at the DSMZ-German Collection of Microorganisms and Cell Cultures (DSMZ, Leibniz Institute, Braunschweig, Germany) under reference ACC-439. G292, HOS, 143B, MG63, SaOS2, and CAL-72 cell lines were cultured in DMEM (Lonza) and the SJSA-1 cell line in RPMI-1640 (Lonza), both supplemented with 10% FBS.
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