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Saos 2

Manufactured by Procell
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Sourced in China

Saos-2 is a human osteosarcoma cell line. It is a well-established model for the study of osteoblast-like cells and bone-related research.

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Lab products found in correlation

Hfob1, by Procell (8 mentions) Sw1353, by Thermo Fisher Scientific (1 mentions) Mccoy s 5a medium, by Procell (1 mentions) Sh circ, by GenePharma (1 mentions) Sw1353, by Procell (1 mentions) Si col1a1, by GenePharma (1 mentions) Si nc, by GenePharma (1 mentions) Fetal bovine serum (fbs), by Procell (1 mentions) Penicillin streptomycin, by Procell (1 mentions) Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions) Goat anti mouse igg, by OriGene (1 mentions) Mccoy s 5a, by Thermo Fisher Scientific (1 mentions) Fibronectin rabbit polyclonal antibody, by Abcam (1 mentions) Transwell chamber, by Corning (1 mentions) Ab32572, by Abcam (1 mentions) Ab32072, by Abcam (1 mentions) Saos 2, by Solarbio (1 mentions) Dulbecco s modified eagle s medium dmem high glucose, by Solarbio (1 mentions) Mccoy s 5a medium, by Thermo Fisher Scientific (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)

13 protocols using saos 2

1

Osteoblast and Osteosarcoma Cell Culture

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Normal osteoblasts (hFOB1.19) and osteosarcoma cells (HOS, Saos-2, and SW1353) were obtained from ProCell (China). SW1353 (Leibovitz’s L-15 Medium; Procell), Saos-2 (McCoy’s 5A; Procell), HOS (MEM; Procell), and hFOB1.19 (DMEM/F12; Procell) were maintained in media containing 1% penicillin-streptomycin (Procell) and 10% fetal bovine serum (FBS, Procell). Culture was performed under a 5% CO2 atmosphere at 37°C.
Small interfering RNAs (siRNAs) targeting circ_0020378 (si-circ_0020378), COL1A1 (si-COL1A1), miR-339-3p inhibitor and mimic, and their respective negative controls (si-NC, inhibitor NC, and mimic NC) were purchased from GenePharma, China. Recombinant lentiviruses (multiplicity of infection [MOI] = 50) carrying short hairpin RNAs (shRNAs) targeting either circ_0020378 (sh-circ) or sh-NC were purchased from GenePharma. Lipo3000 was used to introduce synthetic nucleotides (siRNAs, mimics, and inhibitors) into Saos-2 and HOS cells. After a 48-h transfection period, the efficacy of transfection was verified by RT-qPCR. For shRNA lentivirus infection, approximately 80% confluent Saos-2 cells were incubated with the virus particles (MOI = 50). After 14 days, the survivors were screened by adding 0.5 μg/mL puromycin. Positive monoclonal cells were expanded and verified using RT-qPCR.
Liu L, & Huang W. (2023). hsa_circ_0020378 regulating miR-339-3p/COL1A1 promotes osteosarcoma progression. Cancer Biology & Therapy, 24(1), 2274120.
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2

Culturing Human Osteoblast and Osteosarcoma Cells

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Human osteoblast cell line (HfoB1.19) and human OS cell lines (MG63, U2OS, and Saos-2) were all purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China).
Cell culture: HfoB1.19 cells, MG63, U2OS, and Saos-2 OS cells were cultured in DMEM medium supplemented with 10% fetal bovine serum (FBS). The media were purchased from Procell Life Science & Technology Corporation (Wuhan, China), and the FBS were purchased from Hyclone (South Logan, UT, USA). The medium contained penicillin (100 U/mL) and streptomycin (100 U/mL). All cell lines were grown in a 37°C incubator with 5% CO2.
Liu B., Zhan X, & Liu C. (2021). Long Noncoding RNA MALAT1 Interacts with miR-124-3p to Modulate Osteosarcoma Progression by Targeting SphK1. Journal of Oncology, 2021, 8390165.
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3

Osteosarcoma Cell Lines Characterization

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Osteosarcoma cells Saos-2 (CL-0202), SW1353 (CL-0447), and HOS (iCell-h099) were purchased from Procell Company (Wuhan, China). U-2OS (ZQ0121), MG63 (ZQ0403), and 143B (ZQ0455) were purchased from Zhong Qiao Xin Zhou Biotechnology Company (Shanghai, China). Osteoblast hFOB 1.19 (ZQ0402) was purchased from Zhong Qiao Xin Zhou Biotechnology Company. All of the cell lines were authenticated by short tandem repeat DNA profiling and were found to be free of mycoplasma infection. Saos-2 cells were cultured in McCoy’s 5A medium (PM150710, Procell Company) with 15% fetal bovine serum (FBS) (P10033, Hyclone, USA). SW1353, U-2OS, and 143B cells were cultured in Dulbecco’s modified Eagle medium (DMEM) (12100-46, Gibco, USA) with 10% FBS. MG63 and HOS cells were cultured in minimum Eagle’s medium with 10% FBS, and hFOB 1.19 cells were cultured in DMEM/F12 medium with 10% FBS and 0.3 mg/ml G418. All cells were maintained in the humidified indicators at 37 °C with 5% CO2.
Yuan Q., Yu H., Chen J., Song X, & Sun L. (2020). ADAM10 promotes cell growth, migration, and invasion in osteosarcoma via regulating E-cadherin/β-catenin signaling pathway and is regulated by miR-122-5p. Cancer Cell International, 20, 99.
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4

Silencing PTN in Osteoblast and OSA Cells

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The osteoblast cell line (hFOB1.19) and human OSA cell lines (HOS, Saos-2 and 143B) were purchased from the Procell Life Science&Technology Co.,Ltd. (Wuhan, China). The cell lines were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin and were grown in an incubator at 37°C with 5% CO2. The silencing RNA against PTN (si-PTN) was synthesized and purchased from TsingkeBiotechnology Co.,Ltd. (Beijing, China). The sequence of si-PTN is shown in Supplementary Table 1. Lipofectamine 3000 (Invitrogen) was used for cell transfection.
Jia C., Liu M., Yao L., Zhao F., Liu S., Li Z, & Han Y. (2024). Multi-omics analysis reveals cuproptosis and mitochondria-based signature for assessing prognosis and immune landscape in osteosarcoma. Frontiers in Immunology, 14, 1280945.
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5

Osteosarcoma Tissue and Cell Line Analysis

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This study was approved by the ethics committee of the General Hospital of Ningxia Medical University (No. KYLL-2021-562), and a written informed consent form was signed by all patients. Ten osteosarcoma samples and matched adjacent normal tissues were collected from August 2021 to August 2022, and postoperative pathology indicated osteosarcoma. The specimens were frozen and stored at -80°C until use.
Osteosarcoma cell lines (U-2OS, MG-63, and Saos-2) and a normal osteoblast cell line (hFOB 1.19) were purchased from Procell Life Science & Technology (Wuhan, Hubei, China), and verified using STR genotyping. U-2OS and Saos-2 cells were cultured in McCoy's 5A medium supplemented with 10% FBS (fetal bovine serum) and 1% penicillin and streptomycin. MG-63 cells were grown in minimum essential medium supplemented with 10% FBS and 1% penicillin and streptomycin. hFOB 1.19 cells were cultured in F12 medium and Dulbecco’s modified Eagle medium supplemented with 0.3 mg/ml G418, 10% FBS, and 1% penicillin and streptomycin. hFOB 1.19 cells were maintained in an incubator at 34°C with 5% CO2, and all other cells were maintained at 37°C with 5% CO2.
Wan N., Liu Q., Shi J, & Wang S. (2024). LncRNA SNHG25 Predicts Poor Prognosis and Promotes Progression in Osteosarcoma via the miR-497-5p/SOX4 Axis. Combinatorial Chemistry & High Throughput Screening, 27(5), 725-744.
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6

Investigating Osteosarcoma Cell Signaling

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Osteosarcoma cell lines (SAOS2, MG63 and U2OS) and the human osteoblast cell line (hFOB1.19) were purchased from Procell Co. Ltd (Wuhan, China). PRR11 rabbit anti-human polyclonal antibody (Ab237526), β-catenin rabbit anti-human monoclonal antibody (ab32572), p-β-catenin rabbit anti-human monoclonal antibody (ab75777), GSK-3β mouse anti-human monoclonal antibody (ab93926), p-GSK-3β rabbit anti-human monoclonal antibody (ab68476), c-Myc rabbit anti-human monoclonal antibody (ab32072), CyclinD1 rabbit anti-human monoclonal antibody (ab16663), E-cadherin mouse anti-human monoclonal antibody (Ab1416), Vimentin rabbit anti-human monoclonal antibody (ab92547), Fibronectin rabbit polyclonal antibody (ab2413), total RNA extraction kit were purchased from Abcam (USA). McCoy's 5A and 10% fetal bovine serum-containing DEME were purchased from GIBCO (Thermo Fisher Scientific, USA). Goat anti-mouse IgG, goat anti-rabbit IgG, DAB color kit was purchased from OriGene (USA). The reverse transcription kit HiScript reverse transcriptase and SYBR Green Master Mix were purchased from Novozan Biotechnology Co., Ltd (Nanjing, China). Transwell chamber were purchased from Corning, (USA), and CCK8 kit was purchased from MCE, (USA). PRR11 targeting SiRNA was purchased from Jima (Shanghai, China).
Li K., Yu H., Zhao C., Li J., Tan R, & Chen L. (2021). Down-regulation of PRR11 affects the proliferation, migration and invasion of osteosarcoma by inhibiting the Wnt/β-catenin pathway. Journal of Cancer, 12(22), 6656-6664.
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7

Osteosarcoma and Osteoblast Cell Culture

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Human osteosarcoma cells (Saos2, 143B) and normal osteoblast cell line (hFOB 1.19) were provided by the Procell Company (Wuhan, China). Saos2 cells were cultured in McCoy’s medium supplemented with 15% FBS, while 143B cells were in RPMI 1640 medium supplemented with 10% FBS. hFOB cells were cultured in DMEM/F-12 medium. All osteosarcoma cell lines were maintained in a humidified incubator at 37 °C with 5% CO2, while hFOB 1.19 cells were cultured at 35 °C with 5% CO2.
Liu Y., He M., Tang H., Xie T., Lin Y., Liu S., Liang J., Li F., Luo K., Yang M., Teng H., Luo X., He J., Liao S., Huang Q., Feng W., Zhan X, & Wei Q. (2024). Single-cell and spatial transcriptomics reveal metastasis mechanism and microenvironment remodeling of lymph node in osteosarcoma. BMC Medicine, 22, 200.
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8

Establishing Osteosarcoma-Resistant Cell Lines

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Two OS cell lines MG63 and Saos-2 were obtained from Procell (Wuhan, China). Through the stepwise increasing CDDP concentrations (0–60 μg/mL, Sigma, Shanghai, China), the corresponding OS-resistant cell lines (MG63/CDDP and Saos-2/CDDP) were established from parental cell lines MG63 and Saos-2, as previously described.22 (link) All cells were cultivated in Dulbecco’s modified Eagle’s medium (DMEM) high-glucose (4.5g/L) (Solarbio, Beijing, China) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific, Rockville, MD, USA). All cells were incubated in an incubator with 5% CO2 at 37°C.
Liu L, & Wang S. (2020). Long Non-Coding RNA OIP5-AS1 Knockdown Enhances CDDP Sensitivity in Osteosarcoma via miR-377-3p/FOSL2 Axis. OncoTargets and therapy, 13, 3853-3866.
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9

Culturing Human Cell Lines

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Human normal fetal osteoblast (hFOB1.19), OS cell lines (U2OS, HOS and SaoS2) and human umbilical vein endothelial cells (HUVECs) were purchased from Procell (Wuhan, China), and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Weike Biotechnology, Shanghai, China), McCoy’s 5a medium (Gibco, Carlsbad, CA, USA) or Ham’s F12K (Procell) added with fetal bovine serum (FBS; Biosun, Shanghai, China) and 1% penicillin/streptomycin (Gibco). hFOB1.19 cells were cultured at 34 °C, and other cells grew at 37 °C.
Wang W., Wang J., Li Y, & Zhao Y. (2022). Circ_0051079 silencing inhibits the malignant phenotypes of osteosarcoma cells by the TRIM66/Wnt/β-catenin pathway in a miR-625-5p-dependent manner. Journal of Bone Oncology, 35, 100436.
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10

Knockdown of DEPDC1 and Overexpression of KIF4A in Osteosarcoma Cells

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hFOB1.19, U2OS, SaOS-2 and MG-63 cells were obtained from Procell (Wuhan, China). Human umbilical vein endothelial cells (HUVECs) were provided from American Type Culture Collection (ATCC). All cells were cultivated within the Dulbecco’s Modified Eagle Medium (DMEM) containing 10% fetal bovine serum (FBS) at 37 °C in an incubator under 5% CO2.
To knock down DEPDC1 expression and overexpress KIF4A expression in osteosarcoma cells, short hairpin (sh)RNA targeting ANGPT2 (sh-DEPDC1#1 and sh-DEPDC1#2), pcDNA3.1-KIF4A as well as corresponding negative control (sh-NC) and pcDNA3.1-NC were obtained from RIBIO (Guangzhou, China). When OS cells were reached to about 80% confluence, OS cells were transfected with different vectors using Lipofectamine 3000 according to the manufacturer’s protocol. The transfection efficiency was confirmed by RT-qPCR and western blot 48 h later.
Yang M., Zhang H., Gao S, & Huang W. (2023). DEPDC1 and KIF4A synergistically inhibit the malignant biological behavior of osteosarcoma cells through Hippo signaling pathway. Journal of Orthopaedic Surgery and Research, 18, 145.
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