Snu 182

Manufactured by Thermo Fisher Scientific

Sourced in United States

The SNU-182 is a laboratory instrument designed for general use in scientific research and analysis. It is a compact, benchtop device that performs specific functions related to sample handling and processing. The core functionality of the SNU-182 is to facilitate various laboratory procedures, but a more detailed description cannot be provided while maintaining an unbiased and factual approach.

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11 protocols using snu 182

Generating Stable FOXF1-Overexpressing Cell Lines

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Five human HCC cell lines HepG2, SMMC-7221,SNU-182, Hep3B, and SNU-449 were obtained from the Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, People’s Republic of China), and were cultured in Roswell Park Memorial Institute 1640 medium (Thermo Fisher Scientific) containing 10% fetal bovine serum (FBS) (Thermo Fisher Scientific) at 37°C with 5% CO₂.
To get a stable FOXF1-overexpressing cell line, the FOXF1 expression plasmid pCDNA3.1-FOXF1 was transfected into SNU-182 cells, followed by selection on 800 μg/mL of neomycin for 21 days. FOXF1-specific siRNA was transfected into HepG2 cells. All the transfection experiments used Lipofectamine 2000 (Thermo Fisher Scientific), and the cells were incubated according to the manufacturer’s instruction.

Zhao Z.G., Wang D.Q., Hu D.F., Li Y.S, & Liu S.H. (2016). Decreased FOXF1 promotes hepatocellular carcinoma tumorigenesis, invasion, and stemness and is associated with poor clinical outcome. OncoTargets and therapy, 9, 1743-1752.

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Characterization of Human Liver Cell Lines

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Human liver epithelial cells (THLE-2, CBP61031) and the human liver cancer cell lines, SNU-182 (CBP60211), SNU-387 (CBP60214), Hep3B (CBP60197) and PLC/PRF/5 (CBP60223), were purchased from Nanjing Cobioer BioScience Co., Ltd. The human liver cancer cell line, SK-Hep1 (HTB-52), was obtained from the American Tissue Culture Collection (ATCC). THLE-2 cells were cultured in Dulbecco's modified Eagle's medium (DMEM, 10566024; Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS, 16140071; Gibco; Thermo Fisher Scientific, Inc.), 100 IU/ml penicillin and 100 µg/ml streptomycin (15070063; Gibco; Thermo Fisher Scientific, Inc.). The SNU-182, SNU-387 and PLC/PRF/5 cells were cultured in RPMI-1640 medium (61870044; Gibco; Thermo Fisher Scientific, Inc.) with 10% heat-inactivated FBS in 5% CO₂. The SK-Hep1 and Hep3B were grown in DMEM supplemented with 10% FBS, 2 mM L-glutamine (4 mM, 25030081; Gibco; Thermo Fisher Scientific, Inc.), penicillin and streptomycin.

Hu Z., Zhang Z., Teng F., Feng J., Wu X, & Chang Q. (2020). Role of Asxl2 in non-alcoholic steatohepatitis-related hepatocellular carcinoma developed from diabetes. International Journal of Molecular Medicine, 47(1), 101-112.

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Cancer Cell Line Culture Protocols

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The human colorectal cancer cell (CRC) lines HCT-116 and HT-29, human ovarian cancer cell lines Caov-3 and SK-OV-3, human breast cancer MCF7, MDA-MB-361 and SK-BR-3, human lung cancer A549 and NCI-H460, human gastric cancer AGS and NCI-N87, human liver cancer HEPG2 and SNU-182 cell lines were purchased from the American Type Culture Collection (ATCC). HCT-116, HT-29, SK-OV-3, and SK-BR-3 cells were cultured in McCoy’s 5A medium (Thermo Fisher Scientific). MCF7 and HEPG2 cells were cultured in Eagle’s Minimum Essential Medium (Thermo Fisher Scientific). Caov-3 cell was cultured in Dulbecco’s Modified Eagle’s Medium (Thermo Fisher Scientific). MDA-MB-361 cell was cultured in Leibovitz’s L-15 medium (Thermo Fisher Scientific). A549 and AGS cells were cultured in F-12 K medium (Thermo Fisher Scientific). NCI-H460, NCI-N87, and SNU-182 cells were cultured in RPMI-1640 medium (Thermo Fisher Scientific). The complete growth medium was produced by the addition of 1 % penicillin-streptomycin (Thermo Fisher Scientific) and 10 % fetal calf serum (Thermo Fisher Scientific). The e-Myco VALiD Mycoplasma PCR Detection Kit (iNtRon Biotechnology) was used to confirm the absence of mycoplasma. All cell lines used in this study were examined by short tandem repeat profiling.

Liu J., Qiu J., Zhang Z., Zhou L., Li Y., Ding D., Zhang Y., Zou D., Wang D., Zhou Q, & Lang T. (2021). SOX4 maintains the stemness of cancer cells via transcriptionally enhancing HDAC1 revealed by comparative proteomics study. Cell & Bioscience, 11, 23.

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Cell Culture Protocols for Liver Cancer

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The HCC cell lines SNU-182, Huh7, and Hep 3 B, and the normal liver cell line THLE2 (control) were obtained from Procell (Wuhan, China) and BeNa Co., Ltd (Beijing, China). Huh7 and Hep 3 B cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM; GIBCO, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS; GIBCO), SNU-182 cells were maintained in Roswell Park Memorial Institute (RPMI) 1640 medium (GIBCO) supplemented with 10% FBS, and THLE2 cells were maintained in FBS-containing (10%) bronchial epithelial growth medium (BEGM; GIBCO). Cell cultures were maintained at 37°C in a humidified incubator with 5% CO₂. Cells at 80% confluency were passaged, and cells at the logarithmic phase were used in subsequent experiments.

Chen Z., Qi L., Fu H, & Ma L. (2022). Long non-coding RNA X-inactive specific transcript suppresses the progression of hepatocellular carcinoma through microRNA-221-3p-targeted regulation of O6-methylguanine-DNA methyltransferase. Bioengineered, 13(5), 14013-14027.

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Cultivation of Hepatocellular Carcinoma Cell Lines

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HCC cell lines SNU-449, SNU-182, Huh7, LM3, Bel-7405, SK-hep1, Hep3B and normal human liver cell line L02 were purchased from the American Type Culture Collection (Manassas, VA, USA) or Institute of Biochemistry and Cell Biology (Chinese Academy of Sciences, Shanghai, China). SNU-449, SK-hep1 and SNU-182 were cultured in RPMI-1640 medium (Gibco, USA) containing 10% fetal bovine serum (FBS, Gibco). The other cells were cultured in high-glucose Dulbecco’s modified Eagle medium (DMEM) (Gibco) containing 10% FBS. Penicillin (100 U/ml) and streptomycin (100 μg/ml) were supplemented to the medium to reduce the chance of microbial contamination. All the cells were maintained in a humidified atmosphere at 37 °C with 5% CO₂/95% air.

Shu G., Su H., Wang Z., Lai S., Wang Y., Liu X., Dai L., Bi Y., Chen W., Huang W., Zhou Z., He S., Dai H, & Tang B. (2021). LINC00680 enhances hepatocellular carcinoma stemness behavior and chemoresistance by sponging miR-568 to upregulate AKT3. Journal of Experimental & Clinical Cancer Research : CR, 40, 45.

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Cell Line Source and Culture Conditions

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THLE-2, MIHA, Hep3B and Huh7 cells were purchased from the National Collection of Authenticated Cell Cultures (https://www.cellbank.org.cn/). SNU-449, SNU-182, and SNU-387 were purchased from ATCC: The Global Bioresource Center (ATCC, https://www.atcc.org/). PLC/PRF/5 and MHCC-97H were provided by Prof. Lu Guo-dong of the School of Public Health, Guangxi Medical University. Hep3B was cultured in MEM medium (Gibco, Shanghai, China) supplemented with 10% FBS (Gibco, Shanghai, China). THLE-2 was cultured in BEGM medium (Lonza, CC3170, Hong Kong). SNU-449, SNU-182, and SNU-387 were cultured in RMPI-1640 medium (Gibco, Shanghai, China) supplemented with 10% FBS (Gibco, Shanghai, China). MIHA, LM3, Huh7, PLC5, and MHCC97-H were cultured in DMEM medium (Gibco, Shanghai, China) supplemented with 10% FBS (Gibco, Shanghai, China).

Zhou X., Luo J., Xie H., Wei Z., Li T., Liu J., Liao X., Zhu G, & Peng T. (2022). MCM2 promotes the stemness and sorafenib resistance of hepatocellular carcinoma cells via hippo signaling. Cell Death Discovery, 8, 418.

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Maintaining Human Cancer Cell Lines

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HEK 293T cell line and human liver cancer cell lines (Huh-7, SNU-449, SNU-182, SNU-387, PLC/PRF/5 and Hep3B) were obtained from ATCC and stored in our laboratory. HEK 293T, Hep3B and Huh-7 cells were maintained in Dulbecco's modified Eagle Medium supplemented with 10% foetal bovine serum (Gibco, Carlsbad, CA, USA). SNU-449, SNU-182, SNU-387, SMMC-7721 and PLC/PRF/5 cells were maintained in RPMI-1640 medium supplemented with 10% foetal bovine serum (Gibco). All cell lines were maintained in a humidified incubator containing 5% CO₂ at 37 °C.

Lv J., Zhu P., Yang Z., Li M., Zhang X., Cheng J., Chen X, & Lu F. (2014). PCDH20 functions as a tumour-suppressor gene through antagonizing the Wnt/β-catenin signalling pathway in hepatocellular carcinoma. Journal of Viral Hepatitis, 22(2), 201-211.

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Molecular Profiling of Liver Hepatocellular Carcinoma

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Thirty-five LIHC specimens and matched paracancerous normal tissues were obtained from patients diagnosed with LIHC at our hospital. All patients provided informed consent, and the ethics committee of the hospital approved our study. Patient characteristics are shown in Table 1. LIHC cell lines (Hep3B, HCCLM3, Huh7, and SNU-182) and normal liver epithelial cells (THLE2) were provided by American Type Culture Collection (ATCC, USA). Hep3B, HCCLM3, and Huh7 cells were cultured in DMEM (Gibco, USA). SNU-182 and THLE2 cells were cultured in medium RPMI-1640 (Gibco). All cells were incubated with 10% FBS at 37 °C and 5% CO₂. siRNA targeting lncRNA ST8SIA6-AS1 or HMGA1 (si-lnc or si-HMGA1), miR-142-3p mimics/inhibitor (anti-miR), and their negative controls (NC) (RiboBio Co. Ltd., China) were used to treat HCCLM3 and Huh7 cells using Lipofectamine 2000 (Invitrogen, USA). After 48 h, further functional analyses were performed.Table 1

Baseline characteristics of 35 HCC patients.

Categories	Cases (Total n = 35)	Percentage (%)
Gender
Male	14	40.0
Female	21	60.0
Age (years)
≥ 55	18	51.4
< 55	17	48.6
Tumor size (cm)
≥ 5	11	31.4
< 5	24	68.6
TNM stage
I + II	20	57.1
III + IV	15	42.9
Lymph node metastasis
Positive	19	54.3
Negative	16	45.7
Liver cirrhosis
Presence	13	37.1
Absence	22	62.9
Serum AFP (ng/mL)
≤ 20	18	51.4
> 20	17	48.6

Feng T., Yao Y., Luo L., Zou H., Xiang G., Wei L., Yang Q., Shi Y., Huang X, & Lai C. (2023). ST8SIA6-AS1 contributes to hepatocellular carcinoma progression by targeting miR-142-3p/HMGA1 axis. Scientific Reports, 13, 650.

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Culturing and Transfecting Human Liver Cell Lines

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Human HCC cell lines Hep 3B (HB-8064), HEP G2 (HB-8065), SNU-182 (CRL-2235), SNU-387 (CRL-2237), human normal liver cell line THLE-3 (CRL-11233) and the HEK293t cell line used for dual-luciferase reporter assay were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Gibco, NY) containing 10% fetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin. Cells were placed in an incubator at 37°C with 5% CO_2. Above cell lines and medium were purchased from American Type Culture Collection (ATCC; Manassas, VA, USA).
For cell transfection, HCC cells were seeded onto 6-well plates with a density of 2×10⁵ cells/well. When the cell confluence reached to 80%, miR-424-5p mimic (miR-mimic) or corresponding control (NC-mimic) and E2F7 overexpression plasmids (oe-E2F7) or empty vector control (oe-NC) were respectively transiently transfected into target cells using lipofectamin2000 (Invitrogen, Carlsbad, USA) according to instructions. Medium was replaced after 6 h and cells were collected after 48 h of transfection. miR-mimic and NC-mimic were purchased from GenePharma (Shanghai, China), while oe-E2F7 and oe-NC were purchased from RiboBio company (Guangzhou, China).

Zhao Y., Zhu C., Chang Q., Peng P., Yang J., Liu C., Liu Y., Chen X., Liu Y., Cheng R., Wu Y., Wu X., Hu L, & Yin J. (2020). MiR-424-5p regulates cell cycle and inhibits proliferation of hepatocellular carcinoma cells by targeting E2F7. PLoS ONE, 15(11), e0242179.

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Establishment and Characterization of HCC Cell Lines

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A total of 74 pairs of HCC cancerous and corresponding non-cancerous tissues were gathered from HCC patients who underwent surgery from 2017 to 2020 at The First A liated Hospital of Guangxi Medical University. These tissue samples were immediately transferred into liquid nitrogen right after the surgery and then stored at -80 ºC for future use. This study was reviewed and approved by the ethics committee of The First A liated Hospital of Guangxi Medical University, and written informed consent in accordance with the Declaration of Helsinki and its later revision was provided by all the patients.
Cell lines and cell culture HCC cell lines SNU-449, SNU-182, Huh7, LM3, Bel-7405, SK-hep1, Hep3B and normal human liver cell line L02 were purchased from the American Type Culture Collection (Manassas, VA, USA) or Institute of Biochemistry and Cell Biology (Chinese Academy of Sciences, Shanghai, China). SNU-449, SK-hep1 and SNU-182 were cultured in RPMI-1640 medium (Gibco, USA) containing 10% fetal bovine serum (FBS, Gibco). The other cells were cultured in high-glucose Dulbecco's modi ed Eagle medium (DMEM) (Gibco) containing 10% FBS. Penicillin (100 U/ml) and streptomycin (100 μg/ml) were supplemented to the medium to reduce the chance of microbial contamination. All the cells were maintained in a humidi ed atmosphere at 37℃ with 5% CO 2 /95% air.

Shu G., Su H., Wang Z., Lai S., Wang Y., Liu X., Dai L., Bi Y., Chen W., Huang W., Zhou Z., He S., Dai H., & Tang B. (2021). LINC00680 enhances hepatocellular carcinoma stemness behavior and chemoresistance by sponging miR-568 to upregulate AKT3.

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