Snu 398

Manufactured by Thermo Fisher Scientific

Sourced in United States, China

The SNU-398 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed for use in scientific research and analysis. The core function of the SNU-398 is to provide a controlled environment for sample preparation and processing. Further details about the specific features and intended applications of this product are not available.

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16 protocols using snu 398

Hepatocellular Carcinoma Cell Lines: Knockdown of ALG3

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The human liver cell line LO2 and hepatocellular cell lines SNU398, SNU449, MHCC97H, Hep3B, PLC/PRF/5, and HepG2 were purchased from the ATCC (Manassas, VA, United States). PLC/PRF/5, Hep3B, MHCC97H, and HepG2 cell lines were cultured in complete Dulbecco’s modified Eagle’s medium (Gibco, United States), and SNU398 and SNU449 cells were cultured in RPMI1640 medium (Gibco, United States); all cells were supplemented with 10% fetal bovine serum (FBS) at 37°C in a humidified incubator with 5% CO₂ in the air.
HCC cell lines (Hep3B, HepG2, and SNU398) were transfected with siRNAs targeting ALG3 or control siRNA (HIPPOBIO, China) using Lipofectamine 2000 Reagent (Invitrogen, United States). The efficiency of the knockdown was confirmed using qRT-PCR analysis. The siRNA used were as follows: si-NC: 5′-UUCUUCGAAGGUGUCACGUTT-3′, ALG3 siRNA#1: 5′-GGUUUCGUGUACAUCUUUAUG-3′, and ALG3 siRNA#2: 5′-GGACCUGAGUCUACCCUCAGG-3’.

Zhao Z., Zheng Z., Huang J., Wang J., Peng T., Lin Y, & Jian Z. (2022). Expression of ALG3 in Hepatocellular Carcinoma and Its Clinical Implication. Frontiers in Molecular Biosciences, 9, 816102.

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Isolation and Transfection of HCC Cells

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Fine needle aspiration was performed to obtain HCC and paired non-tumor tissues from the 62 HCC patients. After histopathological con rmation, all tissue samples were immediately subjected to RNA isolation.
HCC cell line SNU-398 (ATCC, USA) was included. RPMI 1640 (90%) was mixed with FBS (10%) to serve as culture medium for SNU-398 cells. In a 5% CO 2 and 95% humidity incubator cells were cultivated at 37 °C.
Transient transfections DLGAP1-AS2 siRNA and negative control (NC) siRNA, as well as NC miRNA and mimic of miR-154-5p were all purchased from Invitrogen (Shanghai, China). Through lipofectamine 2000 (Invitrogen)-mediated transient transfections, SNU-398 cells were transfected with 40 nM siRNA or miRNA. To perform NC experiments, cells were transfected with either NC siRNA or NC miRNA. Incubation with transfection mixture was performed for 6h, followed by cell culture in fresh medium for further 48h. To perform Control (C) experiments, untransfected cells were cultivated in fresh medium for 54h.
RNA isolation and process SNU-398 cells and paired tissue samples were subjected to RNA isolations, which were performed using RNAzol (Invitrogen). Incubation with DNase I (Invitrogen) was performed for 100 min at 37°C to achieve complete genomic DNA removal. Ratios of OD260/280 were determined to check RNA purity.

Chen K., Zhang Z., Yu A., Li J., Liu J., & Zhang X. (2020). LncRNA DLGAP1-AS2 SiRNA Silencing Inhibits Hepatocellular Carcinoma Cell Migration and Invasion by Up-regulating MiR-154-5p Through Methylation.

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Comparative Cytotoxicity Assay Protocol

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DAPT and gliotoxin, were obtained from Merck Millipore (Darmstadt, Germany). The compounds were reconstituted in dimethyl sulfoxide (DMSO). Human cell lines derived from melanoma (518A2), HCC (HEP3B, SNU398, Huh7), pancreas-CA (PANC1), and breast-CA (HCC38, MDA-MB-468) were cultured in RPMI 1640 supplemented with 10% heat inactivated fetal calf serum (FCS), 2 mM Glutamine, 100 U/ml penicillin, and 100 mg/ml streptomycin (all reagents were obtained from Gibco, Life Technologies Inc., Paisley, United Kingdom). HEP3B (HB-8064), SNU398 (CRL-2233), PANC1 (CRL-1469), HCC38 (CRL-2314), and MDA-MB-468 (HTB-132) cell lines were obtained from the American Type Culture Collection (ATCC, Rockville, MA, United States). The cell line Huh7 (JCRBO 403) was obtained from the National Institute of Biomedical Innovation (Osaka, Japan). The melanoma cell line 518A2, characterized by the BRAF V600E mutation and a CDKN2A exon 2 deletion, was obtained from Leiden University. The generation of the HCC cell line HCC-3 was described previously (Winter et al., 2008 (link)). Cells were incubated with the indicated concentrations of inhibitors or with equal amounts of solvent.

Hubmann R., Sieghart W., Schnabl S., Araghi M., Hilgarth M., Reiter M., Demirtas D., Valent P., Zielinski C., Jäger U, & Shehata M. (2017). Gliotoxin Targets Nuclear NOTCH2 in Human Solid Tumor Derived Cell Lines In Vitro and Inhibits Melanoma Growth in Xenograft Mouse Model. Frontiers in Pharmacology, 8, 319.

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Silencing G6PD in Liver Cancer Cells

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We purchased WRL68 liver normal cells from Shanghai Fuheng Biotechnology Co., LTD. (Fuheng, Shanghai, China), LI-7, SNU-398, SNU-449, SK-HEP-1 Liver cancer cells were acquired from Guangzhou Cellcook Biotechnology Co., LTD (Cellcook, Guangzhou, China). Dulbecco’s modified Eagle’s medium (DMEM; Gibco, Shanghai, China) was used to culture WRL68. RPMI 1640’s modified Eagle’s medium was used to culture SNU-398 (RPMI 1640 modified; Gibco, Shanghai, China). Meanwhile, we cultured LI-7, SNU-449, SK-HEP-1 cells using RPMI 1640 medium. siRNA-G6PD and non-targeting control siRNA (NC-siRNA) were purchased from Ribobio Biotechnology Co., LTD. (Ribobio, Guangzhou, China). The sequence of siRNA-G6PD was TCCTCTATGTGGAGAATGA. The stareffect II transfection reagent (GenStar, Beijing, China) was bound to siRNA-G6PD or NC-SIRRNA for 10 min and transfected into LI-7 or SNU-449 cells. The solution was changed after 6 h, and cells were collected after 48 h.

Li F., Wang B., Li H., Kong L, & Zhu B. (2024). G6PD and machine learning algorithms as prognostic and diagnostic indicators of liver hepatocellular carcinoma. BMC Cancer, 24, 157.

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Hepatic Cell Lines Culture Conditions

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Human hepatic L02 cell line, HCC cell lines MHCC-97 h, MHCC-97 L were provided by the Cell Bank of the Chinese Academy of Sciences (Shanghai, China), SNU-398, PLC/PRF/5 were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA), HLE, Huh-7 were obtained from the Japanese Cancer Research Bank (Tokyo, Japan). SNU-398 were cultured in RPMI 1640 Medium (GIBCO), the rest of the cell lines were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; GIBCO, Grand Island, NY, USA). All medium was supplemented with 10% fetal bovine serum (FBS; GIBCO) and 1% penicillin/streptomycin (Invitrogen). Cells were cultured in a humidified atmosphere of 5% CO₂ at 37 °C. The Notch signaling pathway inhibitor RO4929097 was purchased from MedChemExpress (MCE).

Wang X., Wang R., Bai S., Xiong S., Li Y., Liu M., Zhao Z., Wang Y., Zhao Y., Chen W., Billiar T.R, & Cheng B. (2019). Musashi2 contributes to the maintenance of CD44v6+ liver cancer stem cells via notch1 signaling pathway. Journal of Experimental & Clinical Cancer Research : CR, 38, 505.

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Cell Lines for Hepatocellular Carcinoma Research

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Human HCC cell line SNU398, SNU449, SK-Hep-1, Hep3B, HepG2/C3A, as well as Human Embryonic Kidney 293T (HEK-293T) cell line were purchased from the American Type Culture Collection (ATCC, USA). Human HCC cell lines SMMC-7721, Huh7, and normal human hepatocyte cell line LO2 were purchased from the Chinese Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Hep3B cell line was cultured in Eagle's Minimum Essential Medium (MEM; HyClone, USA). SNU398, SNU449 cell line was cultured in RPMI-1640 (Gibco, USA). SMMC-7721, Huh7, SK-Hep-1, HepG2/C3A, LO2, and HEK-293T cell lines were cultured in Dulbecco's Modified Eagle Medium (DMEM; Gibco, USA). During the process of cell culture, we sterilized the experimental materials, and the experiments were performed according to strict aseptic operation; 1% penicillin and streptomycin (Gibco, USA) were added to the growth medium when necessary in an effort to prevent bacterial contamination in the medium. All of the cultured mediums contained 10% (v/v) fetal bovine serum (FBS; Gibco, USA). Cells were cultured in an atmosphere of 95% humidified air and 5% CO₂ at 37 °C.

Xiong J., Lai Y., Cheng N., Liu J., Wang F., Zheng X., Wang Y., Zhuang Q., Lin Y., Liu J., Yang Y., Zhao B, & Yang X. (2023). Lnc-PLA2G4A-4 facilitates the progression of hepatocellular carcinoma by inducing versican expression via sponging miR-23b-3p. Heliyon, 9(8), e18698.

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Cell Line Cultivation and Authentication

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SNU-398, SNU-449, and HEK293T cell lines were purchased from the American Type Culture Collection (ATCC). Huh-7 was purchased from Japanese Collection of Research Bioresources Cell Bank (JCRB). SNU-398 and SNU-449 cells were cultured in Roswell Park Memorial Institute (RPMI) medium (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) (Gibco BRL). Huh-7, SMMC-7721 and HEK293T cells were maintained in Dulbecco's Modified Eagle's medium (DMEM) supplemented with 10% FBS. All cell lines used in this study were regularly authenticated by morphological observation and tested for mycoplasma contamination (MycoAlert, Lonza Rockland, ME, USA). The cells were incubated at 37°C in a humidified incubator containing 5% CO₂.

Qi L., Song Y., Chan T.H., Yang H., Lin C.H., Tay D.J., Hong H., Tang S.J., Tan K.T., Huang X.X., Lin J.S., Ng V.H., Maury J.J., Tenen D.G, & Chen L. (2017). An RNA editing/dsRNA binding-independent gene regulatory mechanism of ADARs and its clinical implication in cancer. Nucleic Acids Research, 45(18), 10436-10451.

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Culturing Hepatocellular Carcinoma Cell Lines

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Transformed Human Liver Epithelial-3 cells (THLE-3) and HCC cell lines (SNU-182, HuH7 and Hep3B) were acquired from the Cell Bank of the Chinese Academy of Sciences. THLE-3 cells were cultured in Bronchial Ep ithelial Growth Medium (Clonetics Corporation) that was supplemented with 10% heat-inactivated FBS (Gibco; Thermo Fisher Scientific, Inc.), 5 ng/ml epidermal growth factor and 70 ng/ml Phosphoethanolamine. The HCC cell line SNU-398 (American Type Culture Collection) was cultured in RPMI-1640 (Gibco; Thermo Fisher Scientific, Inc.) containing 10% FBS (Gibco; Thermo Fisher Scientific, Inc.), 100 U/ml penicillin and 100 ng/ml streptomycin (Gibco; Thermo Fisher Scientific, Inc.). HuH7 and SNU-182 cells were cultured in DMEM (Gibco; Thermo Fisher Scientific, Inc.) and RPMI-1640, respectively, which were supplemented with 10% FBS, 1% GlutaMAX, 1% non-essential amino acids, 100 U/ml penicillin and 100 ng/ml streptomycin. Minimal essential media (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS, 1% GlutaMAX, 1% non-essential amino acids, 1% sodium pyruvate 100 mM solution, 100 U/ml penicillin and 100 ng/ml streptomycin was used to culture Hep3B cells. All cells were maintained at 37°C in a humidified atmosphere containing 5% CO₂.

Gao W., Chen X., Chi W, & Xue M. (2020). Long non-coding RNA MKLN1-AS aggravates hepatocellular carcinoma progression by functioning as a molecular sponge for miR-654-3p, thereby promoting hepatoma-derived growth factor expression. International Journal of Molecular Medicine, 46(5), 1743-1754.

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Cultivation of Human Liver Tumor Cell Lines

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Human liver tumor cell lines (Hep3B, PLC/PRF5, SK-Hep1, SNU398, SNU449, and SNU878) were obtained from the Korean Cell Line Bank (KCLB) (Seoul, Korea). HepG2 cell line was supplied from the American Type Culture Collection (Manassas, VA, USA). Hep3B, SK-Hep1, and HepG2 cells were maintained in a growth medium containing Dulbecco's modified Eagle's medium (DMEM), 10% fetal bovine serum (FBS), and 1% penicillin-streptomycin whereas PLC/PRF5, SNU398, SNU449, and SNU878 cells were cultured in RPMI-1640 media (Gibco, Gaithersburg, USA) containing 10% FBS and 1% penicillin-streptomycin at 37°C in a humidified atmosphere containing 5% CO₂. For all experiments, cells were grown to 80–90% confluence at passages between 10 and 30, and were deprived of serum for 16 h before PPIX treatment.

Lee J.M., Heo M.J., Lee C.G., Yang Y.M, & Kim S.G. (2015). Increase of miR-199a-5p by protoporphyrin IX, a photocatalyzer, directly inhibits E2F3, sensitizing mesenchymal tumor cells to anti-cancer agents. Oncotarget, 6(6), 3918-3931.

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Cultivation of Hepatocellular Carcinoma Cell Lines

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Human immortalized liver cell-line THLE-2 (cat. no. CRL-2706) and HCC cell line SNU-398 (cat. no. CRL-2233) were acquired from American Type Culture Collection (ATCC, Manassas, VA, USA). Human HCC cell lines HuH-7 (cat. no. SCSP-526) and SK-HEP-1 (cat. no. TCHu109) were acquired from the Chinese Academy of Sciences Cell Bank (Shanghai, China). THLE-2 was maintained in the BEGM Bullet Kit (cat. no. CC-3170, Lonza). SNU-398 was maintained in RPMI 1640 medium (Invitrogen Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Invitrogen). HuH-7 was cultured in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10% FBS. SK-HEP-1 was maintained in Eagle’s Minimum Essential Medium (Invitrogen) supplemented with 10% FBS. All cells were cultured at 37°C containing 5% CO₂ and routinely tested as mycoplasma-free.

Tan C., Huang Y., Huang Z., Ning Y., Huang L., Wu X., Lu Y., Wei H, & Pu J. (2023). N6-Methyladenosine-Modified ATP8B1-AS1 Exerts Oncogenic Roles in Hepatocellular Carcinoma via Epigenetically Activating MYC. Journal of Hepatocellular Carcinoma, 10, 1479-1495.

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