The human osteosarcoma cell lines, U2OS (wild-type P53) and SaOS2 (P53-null) were purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea) and cultured in DMEM medium supplemented with 10% fetal bovine serum (Gibco BRL, Gaithersburg, MD), penicillin and streptomycin (100 U/ml), and fungizone (0.25 μg/ml, Gibco BRL, Gaithersburg, MD) at 37 °C in humidified incubator with 5% CO2.
Saos2
Manufactured by Korean Cell Line Bank
SaOS2 is a human osteosarcoma cell line derived from a primary osteosarcoma of the tibia of an 11-year-old Caucasian female. It is an adherent cell line that exhibits an osteoblastic phenotype and is commonly used in research related to bone biology, ossification, and osteosarcoma.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (1 mentions)
Fungizone, by Thermo Fisher Scientific (1 mentions)
Niclosamide, by Cayman Chemical (1 mentions)
Pyrvinium pamoate, by Merck Group (1 mentions)
Sirna duplexes, by Santa Cruz Biotechnology (1 mentions)
Saos2, by Lonza (1 mentions)
Mg 63, by Lonza (1 mentions)
Mp0040, by Merck Group (1 mentions)
Rpmi 1640, by Lonza (1 mentions)
Olaparib, by Santa Cruz Biotechnology (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Lipofectamine rnaimax, by Thermo Fisher Scientific (1 mentions)
5 protocols using saos2
1
Culture of Human Osteosarcoma Cell Lines
2
Osteosarcoma Cell Lines and Reagents
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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.
3
Culturing Human Osteosarcoma Cell Lines
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Human osteosarcoma Saos-2 and U2OS cell lines, obtained from the Korean Cell Line Bank (Seoul, Korea), were maintained at 37°C in a humid atmosphere of 5% CO2 and 95% air in RPMI and DMEM, respectively. Each medium contained 10% FBS and 1% antibiotics.
4
Osteosarcoma Cell Line Cultivation and Manipulation
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The human osteosarcoma cell lines, U2OS, SaOS2, and MG63 were purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea). KHOS/NP cells were kindly provided by Chang-Bae Kong (Department of Orthopedic Surgery, Korea Institute of Radiological and Medical Science) [27 (link)]. The cells were cultured in DMEM media containing 10% FBS (Gibco BRL, Gaithersburg, MD), and 10% penicillin/streptomycin (100 U/mL) at 37 °C in a humidified 5% CO2 incubator. The PARP inhibitor, olaparib, was purchased from Santa Cruz Biotechnology (sc-302017, Santa Cruz Biotechnology, Santa Cruz, CA) and doxorubicin was obtained from Sigma (D1515, Sigma, St. Louis, MO). Short interfering RNA (siRNA) for PARP1 and negative control siRNA duplexes were purchased from Santa Cruz biotechnology (Santa Cruz Biotechnology, Santa Cruz, CA). The cells were transfected using Lipofectamine® RNAiMax (Invitrogen, Life technologies, Carlsbad, CA).
5
Osteoblast Cell Culture on Metallic Coupons
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The human osteoblast cell line, Saos-2, was purchased from the Korean Cell Line Bank (KCBL). Saos-2 cells were cultured in Minimum Essential Medium (MEM, Welgen) supplemented with 10% fetal bovine serum (FBS, #S001-01, Welgen, Republic of Korea) and 1% anti-anti (Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA).
DED, PBF, and TPS test coupons were prepared and cleaned in six steps for in vitro experiments. Firstly, the coupons were sonicated twice in a solution with 1% Solujet (Alconox) at 45 °C. Then, the mixture was sonicated in distilled water twice at 45 °C. After sonication, the coupons were autoclaved at 120 °C for 30 min and dried in a dry oven at 60 °C. Then, each experimental coupon was placed in a 24-well plate for cell culturing. To minimize the number of cells that adhered to the surface of the plastic wells instead of the surface of the coupons, a non-surface-treated 24-well plate (#32024, SPL Life Science) was used and the coupons were designed to fit tightly into the 24-well plate hole. For the positive control, cells were cultured directly on a surface-treated 24-well plate (#30024, SPL Life Science). Saos-2 cells were seeded at a density of 2.4 × 104 cells/well and incubated in a CO2 incubator at 37 °C.
DED, PBF, and TPS test coupons were prepared and cleaned in six steps for in vitro experiments. Firstly, the coupons were sonicated twice in a solution with 1% Solujet (Alconox) at 45 °C. Then, the mixture was sonicated in distilled water twice at 45 °C. After sonication, the coupons were autoclaved at 120 °C for 30 min and dried in a dry oven at 60 °C. Then, each experimental coupon was placed in a 24-well plate for cell culturing. To minimize the number of cells that adhered to the surface of the plastic wells instead of the surface of the coupons, a non-surface-treated 24-well plate (#32024, SPL Life Science) was used and the coupons were designed to fit tightly into the 24-well plate hole. For the positive control, cells were cultured directly on a surface-treated 24-well plate (#30024, SPL Life Science). Saos-2 cells were seeded at a density of 2.4 × 104 cells/well and incubated in a CO2 incubator at 37 °C.
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