Hcclm3

Manufactured by American Type Culture Collection

Sourced in United States, China

The HCCLM3 is a laboratory equipment product. It is a cell culture device designed for the cultivation and maintenance of human cell lines.

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HCCLM3 cells (2 citations) HCCLM3 (LM3) (2 citations)

119 protocols using hcclm3

Silencing PITX2 in Liver Cancer Cells

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Liver cancer cells (Hep3B, MHCC-97H, Huh7, and HCCLM3) and human fetal hepatocyte line (L-02) were purchased from the ATCC (Manassas, VA, USA). Cells were grown in DMEM medium (Gibco BRL, Gaithersburg, MD, USA) containing 10% fetal bovine serum (Gibco BRL).
siRNA targeting PITX2 (si-PITX2-1#: 5′-CAGCCUGAAUAACUUGAACT T-3′ and si-PITX2-2#: 5′-GCCGACTCCTCCGTATGTTTA-3′) and the negative control (si-NC 5′-UUCUCCGAACGUGUCACGUTT-3′) were synthesized in RiboBio (Guangzhou, China). Huh7 was plated in a 96-well plate and transfected with 20 nM siRNAs by Lipofectamine 2000 (Thermo Fisher Scientific, Waltham, MA, USA), and the cells were used for functional assays 2 days post-transfection according to previous research [15 (link)].

Tuerxun K., Zhang S, & Zhang Y. (2021). Downregulation of PITX2 inhibits the proliferation and migration of liver cancer cells and induces cell apoptosis. Open Life Sciences, 16(1), 1322-1329.

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Culturing Human Liver Cell Lines

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Human HCC cell lines (Hep3B, HCCLM3, Huh-7, MHCC97-L) and human normal liver epithelial cell line (THLE-3) purchased from ATCC (Manassas, VA, USA) were allowed to grow in DMEM culture medium (Invitrogen, Carlsbad, CA, USA) with 1% Pen/Strep mixture (Invitrogen) and 10% fetal bovine serum (FBS; Invitrogen). Cell culture was carried out in a humidified atmosphere of 5% CO₂ and 37 °C.

Fa X., Song P., Fu Y., Deng Y, & Liu K. (2021). Long non‐coding RNA VPS9D1-AS1 facilitates cell proliferation, migration and stemness in hepatocellular carcinoma. Cancer Cell International, 21, 131.

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Culturing Diverse Liver Cancer Cell Lines

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The human hepatic normal cell line HL7702 and liver cancer cell lines QGY-7703 and SMMC7721 were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China ), and cultured in high glucose DMEM (HyClone, Logan, USA) containing 10% FBS. The liver cancer cell line HepG2 was purchased from the National Collection of Authenticated Cell Cultures (Beijing, China) and cultured in RPMI 1640 (HyClone) supplemented with 10% FBS (HyClone). Human liver cancer cell lines, including SNU387, Huh7, and HCCLM3 cells, were obtained from ATCC (Manassas, USA) and cultured in high glucose DMEM (HyClone) containing 10% FBS. All cell lines were maintained at 37°C with 5% CO
₂ in a humid atmosphere. Cell lines were tested for mycoplasma contamination using polymerase chain reaction.

Kuang J., Duan T., Gao C., Liu C., Chen S., Zhu L.Y., Min L., Lu C., Wang W, & Zhu L. (2023). RNF8 depletion attenuates hepatocellular carcinoma progression by inhibiting epithelial-mesenchymal transition and enhancing drug sensitivity: The role of RNF8 in EMT and drug sensitivity in HCC. Acta Biochimica et Biophysica Sinica, 55(4), 661-671.

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Culturing Human Liver Cell Lines

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Human HCC cell lines (MHCC97-L, Huh-7, Hep3B and HCCLM3) and the normal human liver immortalized cells (THLE-2) were obtained from ATCC (Rockville, Maryland) and were propagated under 5% CO₂ and 37 °C. DMEM with 10% FBS (Invitrogen) as supplement was available from Invitrogen (Carlsbad, CA) for cell culture.

Min J., Jin D., Zhang F., Kang Y., Qi Y, & Du P. (2021). DLG1-AS1 is activated by MYC and drives the proliferation and migration of hepatocellular carcinoma cells through miR-497-5p/SSRP1 axis. Cancer Cell International, 21, 16.

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Culturing Human Liver Cell Lines

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Human HCC cell lines (HCC-LM3, SK-Hep1, SMMC-7721, SNU182, C3A, HepaG2, Huh7, Hep3B) and human normal liver cell L02 were purchased from the ATCC. All cell lines were cultured in Dulbecco’s modified Eagle medium (DMEM, Gibco, USA) with 10% fetal bovine serum (Gibco, USA), 100 units/mL penicillin, and 100 µg/mL streptomycin (Life Technologies, USA) under 5% CO₂ at 37 °C.

Hua N., Chen A., Yang C., Dong H., He X., Ru G., Tong X., Zhou F, & Wang S. (2022). The correlation of fibrinogen-like protein-1 expression with the progression and prognosis of hepatocellular carcinoma. Molecular Biology Reports, 49(8), 7911-7919.

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Human HCC Cell Line Characterization

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The study conformed to the provisions of the Declaration of Helsinki (as revised in 2013), and was approved by the Ethics Committee of Affiliated Hospital of Nantong University (No. 2022-L094). Informed consent was provided by all participants. Fresh tissue and matched adjacent normal tissue samples from liver cancer patients were surgically removed at the Affiliated Hospital of Nantong University. None of the liver cancer patients received preoperative anti-tumor therapy. The in vitro culture human HCC cell lines (HCCLM3, Huh7, Hep3B) and human normal liver cell line (LO2) were purchased from ATCC (American type culture collection, Manassas, USA).
The cells were cultured in a 37 °C and 5% carbon dioxide (CO₂) environment. The overexpression plasmid, shRNA (short hairpin RNA), and negative control (NC) (Genechem, Shanghai, China) were transfected into cells with Lipofectamine 3000 reagent (Invitrogen, Carlsbad, USA), and the transfection efficiency was evaluated with fluorescent qRT-PCR. The DMEM medium with 10% FBS concentration was used for the cell culture.

Zhang N., Wang F., Zhu L., Chang R., Mok S.R., Peixoto R.D., Tang W, & Chen Z. (2023). Molecular mechanism of the miR-7/BCL2L1/P53 signaling axis regulating the progression of hepatocellular carcinoma. Annals of Translational Medicine, 11(1), 12.

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Exploring METTL3 and USP7 in HCC

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HCC cells (HCCLM3, MHCC97-L, Hep3B, and Huh7) and L02 cell lines (human normal liver cell line) purchased from ATCC (Manassas, VA, USA) were used in this study. METTL3 siRNA, USP7 (ubiquitin-specific protease) siRNA, or negative control together with Lipofectamine 2000 (Beyotime, Shanghai, China) were used for the cell transfection, followed by qRT-PCR for the transfection efficacy determination.

Yuan D., Chen J., Hao Q., Zhang P, & Chen Z. (2022). Methyltransferase-like 3 Aggravates HCC Development via Mediating N6-Methyladenosine of Ubiquitin-Specific Protease 7. Journal of Oncology, 2022, 6167832.

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Culturing Human Liver Cell Lines

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Human hepatoma cell lines (HepG2, MHCC97-H, HCCLM3, SMMC7721 and mhcc97-l) and normal human liver cell lines (LO2) were obtained from ATCC. All cell lines were stored in DMEM (Thermo Fisher Scientific, USA) supplemented with 10% FBS (Thermo Fisher Scientific, USA) at 37° C and 5% CO₂.

Zhang Y., Ni W, & Qin L. (2021). RUFY3 promotes the progression of hepatocellular carcinoma through activating NF-κB-mediated epithelial-mesenchymal transition. Aging (Albany NY), 13(17), 21283-21293.

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Cell Culture of Liver Cancer Lines

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The cell lines used in this study, including LO2, SMMC-7721, Hep3B, HepG2, 97H, 97L, HCC-LM3, and Huh7 were obtained from the ATCC. The cells were cultured in DMEM supplemented with 10% FBS, 100 mg/l streptomycin, and 10⁵ U/L penicillin. The cells were incubated in a 5% CO₂-humidified incubator at 37°C. Liver cancer tissues and the corresponding adjacent healthy tissues were collected from patients treated at Nanjing Drum Tower Hospital between 2018 and 2020. The tissues were stored in liquid nitrogen until use. Before sample collection, written informed consent collection was obtained from all patients.

Xue C., Gao P., Cui X., Zhang X., Lei J., Li R., Zhu C, & Qin X. (2021). ASRGL1 Correlates With Immune Cell Infiltration in Hepatocellular Carcinoma and Can Serve as a Prognostic Biomarker. Frontiers in Oncology, 11, 680070.

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Lentiviral and Plasmid Transfection of Hepatic Cell Lines

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The HCC cell line Hep G2, human normal hepatocyte cell line (L02), and HEK-293 were purchased from ATCC; HCC cell lines SMMC-7721, Huh-7, and HCC-LM3 and human astrocyte cell line LX-2 were purchased from the Institute of Biochemistry and Cell Biology (Chinese Academy of Sciences, Shanghai, China). HCC cells and HEK-293 were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) (Gibco, USA) at 37°C in a humidified atmosphere containing 5% CO₂.
Cell transfections were carried out as described previously [18 (link)]. Briefly, for lentivirus transduction, 10⁵ HCC cells were incubated in a 6-well plate with 2 ml of medium containing 100 μl (10⁷ U) of lentivirus particles and 5 μg/ml polybrene for 24 h. Plasmid and siRNA transfections were performed using Lipofectamine 3000 (Invitrogen, USA) according to the manufacturer's instructions. miRNA mimics or scramble control RNAs (RiboBio, China) were transfected into cells at a final concentration of 100 nM using a riboFECTTM CP Kit (RiboBio, China) according to the manufacturer's instructions.

Chen K., Li Z., Zhang M., Wang B., Peng T., Shen Y., Zhang J., Ye J., Liu Y., Tang D., Peng M., Ma D., Xiao Z., Zhang Y., Jin W, & Li X. (2020). miR-876 Inhibits EMT and Liver Fibrosis via POSTN to Suppress Metastasis in Hepatocellular Carcinoma. BioMed Research International, 2020, 1964219.

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