Huh 7

Manufactured by GE Healthcare

Sourced in United States, China, Austria

The Huh-7 is a laboratory equipment product developed by GE Healthcare. It is a cell line that is widely used in research and drug development. The Huh-7 cell line is derived from human hepatocellular carcinoma. It is a commonly used in vitro model for studying liver-related diseases and for evaluating the efficacy and safety of potential therapeutic agents.

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6 protocols using huh 7

Liver Cancer Cell Line Transfection

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The liver cancer cell lines, HepG2 and Huh-7, were purchased from the American Type Culture Collection (ATCC). The cell lines were authenticated by STR profiling. The cells were maintained in a humidified incubator under standard conditions (37°C, 5% CO₂) in IMDM (PAA Laboratories supplemented with 10% fetal calf serum (FCS).
For siRNA transfection, 5×10² (HepG2) or 3×10² (Huh-7) cells/well were seeded in a 96-well plate, or 3×10⁵ (HepG2) or 2×10⁵ (Huh-7) cells in a 24-well plate, respectively, and incubated under standard conditions unless stated otherwise. Using the INTERFERin™ siRNA transfection reagent (Polyplus), 2.5 nM siRNA (for sequences, see Table SI) for proliferation assays or 5 nM siRNA for all other assays were transfected according to the manufacturer’s instructions and incubated for the time periods as indicated in the figures. The cells were then analyzed in the well plates or harvested by trypsinization and subsequent transfer to Eppendorf cups and used as described below.
Transfection of 400 ng plasmid DNA per 24 well was performed using FuGENE^® transfection reagent (Promega) according to the manufacturer’s protocol and incubated at 37°C for 6 h, prior to media change and siRNA transfection.

Kronschnabl P., Grünweller A., Hartmann R.K., Aigner A, & Weirauch U. (2019). Inhibition of PIM2 in liver cancer decreases tumor cell proliferation in vitro and in vivo primarily through the modulation of cell cycle progression. International Journal of Oncology, 56(2), 448-459.

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Cell Line Culture Conditions

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The cell lines HepG2 and Huh-7 were obtained from American Type Culture Collection (ATCC). The ATP7B knockout derivate of HepG2 (KO) were obtained as described [20 (link)]. HepG2 cells were cultured in RPMI (Lonza). Huh-7 cells were cultured in DMEM High Glucose (GE Healthcare). All media contained 10% fetal bovine serum (FBS) supplemented with 100 U/mL penicillin/streptomycin (Hyclone, Logan). Cell lines were maintained in 5% CO₂ at 37°C in a humidified chamber.

Guttmann S., Chandhok G., Groba S.R., Niemietz C., Sauer V., Gomes A., Ciarimboli G., Karst U., Zibert A, & Schmidt H.H. (2017). Organic cation transporter 3 mediates cisplatin and copper cross-resistance in hepatoma cells. Oncotarget, 9(1), 743-754.

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Cultivation of Various Cell Lines

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The human embryonic kidney cell line 293T and the U87MG human glioma cell line were obtained from the American Type Culture Collection (Manassas, VA, USA). 293FT and Lenti-X 293T cells were purchased from Thermo Fisher Scientific (Waltham, MA, USA) and Takara Bio Inc. (Shiga, Japan), respectively. The A549 human non-small cell lung cancer cell line, PANC-1 human pancreatic cancer cell line, NCI-H1299 human metastatic lung cancer cell line, Huh7 human hepatocellular carcinoma cell line, and CSC2 human glioma stem cell line were obtained from various researchers as described previously^{17 (link)–21 (link)}. The 293T, 293FT, Lenti-X 293T, U87MG, PANC-1, and Huh7 cells were cultured in Dulbecco’s modified Eagle medium (DMEM; HyClone - GE Healthcare, Chicago, IL, USA). The A549 and NCI-H1299 cells were cultured in RPMI 1640 medium (HyClone). The CSC2 glioma stem cells were cultured as described previously^{20 (link)}. All media except that used to culture CSC2 were supplemented with 10% fetal bovine serum (HyClone), 1% penicillin/streptomycin (Welgene, Gyeongsan, Republic of Korea), and 10 μg/ml ciprofloxacin (Santa Cruz Biotech, Santa Cruz, CA, USA).

Seo Y., Kim S.S., Kim N., Cho S., Park J.B, & Kim J.H. (2020). Development of a miRNA-controlled dual-sensing system and its application for targeting miR-21 signaling in tumorigenesis. Experimental & Molecular Medicine, 52(12), 1989-2004.

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Cell Culture Protocol for Cancer and Endothelial Cells

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293T cells and human HCC cell lines Huh7 and Hep3B were purchased from the American Type Culture Collection. Huh7, Hep3B and 293T cells were cultured in Dulbecco's modified Eagles medium (DMEM; HyClone; GE Healthcare Life Sciences) supplemented with 10% fetal bovine serum (HyClone; GE Healthcare Life Sciences), penicillin (100 U/ml) and streptomycin (100 µg/ml) in a humidified atmosphere of 5% CO₂ at 37°C. Human umbilical vein endothelial cells (HUVECs) were purchased from PromoCell Academy and cultured in Endothelial Cell Growth Medium 2 (PromoCell Academy) with Supplement Mix according to the manufacturer's guidelines.

Fan Z., Zheng W., Li H., Wu W., Liu X., Sun Z., Hu H., Du L., Jia Q, & Liu Q. (2020). LOXL2 upregulates hypoxia-inducible factor-1α signaling through Snail-FBP1 axis in hepatocellular carcinoma cells. Oncology Reports, 43(5), 1641-1649.

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Curative Surgical Resection of HCC

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The paired HCC samples and the adjacent non-tumor tissues were collected following curative surgical resection from 48 patients with HCC in the Zhongnan Hospital of Wuhan University between May 2011 and May 2013. This study was approved by the Hospital's Protection of Human Subjects Committee, and written informed consent was obtained from all patients. Based on their medical documents, we conducted a 48-month follow-up survival survey. Overall survival (OS) was defined as the interval between resection and death or the last follow-up visit. Curative resection was defined as the removal of all recognizable tumor tissue with a clear microscopic margin. HepG2, Hep3B, Huh-7, HCCLM9, SK-Hep1, and SMMC-7721 cell lines included in this study were purchased from the Cell Bank of Type Culture Collection (CBTCC, Chinese Academy of Sciences, Shanghai, China) and cultured in Dulbecco's Modified Eagle's Medium (DMEM)/ high-glucose medium (GE Healthcare Life Sciences HyClone Laboratories, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS; Gibco, Thermo Fisher Scientific, Inc., Waltham, MA, USA) and 1% penicillin and streptomycin in a humidified atmosphere of 5% CO 2 at 37°C.

Lan T., Yan X., Li Z., Xu X., Mao Q., Ma W., Hong Z., Chen X, & Yuan Y. (2017). Long non-coding RNA PVT1 serves as a competing endogenous RNA for miR-186-5p to promote the tumorigenesis and metastasis of hepatocellular carcinoma. Tumour biology : the journal of the International Society for Oncodevelopmental Biology and Medicine, 39(6).

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Cellular Expression of Nuclear Receptors

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All cell lines were obtained from the Cell Bank of the Chinese Academy of Science (Shanghai, China). HEK293, Huh7 cell line and normal human hepatic cell line L-02 were maintained in RPMI1640 with 10% FBS (PAA Laboratories, Pasching, Austria). Human HCC cell lines HepG2, Hep3B, SMMC-7721, Huh7 and normal human hepatic cell line Chang were maintained in DMEM with 10% FBS (PAA Laboratories, Pasching, Austria). Antibodies specific to estrogen-related receptor gamma and β-actin were obtained from Sigma (Beijing, China).

Yuan B., Liang Y., Wang D, & Luo F. (2015). MiR-940 inhibits hepatocellular carcinoma growth and correlates with prognosis of hepatocellular carcinoma patients. Cancer Science, 106(7), 819-824.

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