The murine hepatoma cell line Hepa1-6 was purchased from ATCC (CRL-1830; Manassas, VA, United States). GPC3-expressing Hepa1-6 cell line (Hepa1-6/GPC3) was developed according to the established protocol[27 (link)] and cultured in RPMI-1640 supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, and 1 mg/mL G418 (Invitrogen, CA, United States) at 37 oC in a 5% CO2 atmosphere.
Hepa1 6
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Hepa1-6 is a cell line derived from a mouse hepatoma. It is commonly used in research applications that involve the study of liver cell biology and function.
Automatically generated - may contain errors
Lab products found in correlation
Hepa1 6, by American Type Culture Collection (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
Hepg2, by Thermo Fisher Scientific (7 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (5 mentions)
Hep3b, by Thermo Fisher Scientific (4 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Smmc 7721, by Thermo Fisher Scientific (3 mentions)
Hep3b, by American Type Culture Collection (3 mentions)
Dmem medium, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Plc prf 5, by Thermo Fisher Scientific (2 mentions)
Hcclm3, by National Collection of Authenticated Cell Cultures (2 mentions)
L glutamine, by Thermo Fisher Scientific (2 mentions)
Trypan blue, by Beyotime (2 mentions)
Hepa1 6, by National Collection of Authenticated Cell Cultures (2 mentions)
C57bl 6 mice, by American Type Culture Collection (2 mentions)
Ct26 wt, by American Type Culture Collection (2 mentions)
24 protocols using hepa1 6
1
Generation of GPC3-expressing Hepa1-6 Cell Line
2
Culturing HCC Cell Lines for Experiments
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HCC cell lines (SMMC7721, Huh7, HepG2, Hepa1-6) were obtained from the Cell Bank (Academica Sinica, Shanghai, China) and H22-Luc cells were purchased from Shanghai Fuheng Biotechnology. SMMC7721, Huh7, HepG2, and Hepa1-6 were cultured in DMEM (Gibco-Invitrogen) and H22-Luc cells in RPMI 1640 (Gibco-Invitrogen). All culture media were supplemented with 10% FBS (Invitrogen) and 1% penicillin/streptomycin (Invitrogen). Cell lines were cultured in a humidified incubator at 37 °C and 5% CO2.
3
Hepatocyte Cell Lines Establishment
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Hepa 1–6 (murine hepatoma cell lines), Hep3B and HepG2 (human hepatoma cell lines), SKLU-1 (telomerase negative human lung carcinoma cell lines), and HEK293 (adenoviral E1-transformed human embryonic kidney cell lines) were purchased from the American Type Culture Collection (Manassas, VA). Huh7 (human hepatoma cell lines) was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan). Hep3B and HepG2 were cultured in EMEM (Invitrogen, Carlsbad, CA) with 10% FBS. Hepa1–6, SKLU-1, Huh7, and HEK293 cells were maintained in DMEM (Invitrogen) with 10% FBS.
For the establishment of cell lines stably expressing miR-122a dependent on the presence of tetracycline, HepG2 cells expressing Tet repressor vector, pcDNA 6/TR (Invitrogen), were isolated with blasticidin for 2 weeks. The selected cells were transfected with vector encoding pri-miR-122a which was cloned into the tetracycline response element vector, pcDNA 4/myc-His B (Invitrogen), and zeocin resistant colonies were isolated for 2 weeks.
For the establishment of cell lines stably expressing miR-122a dependent on the presence of tetracycline, HepG2 cells expressing Tet repressor vector, pcDNA 6/TR (Invitrogen), were isolated with blasticidin for 2 weeks. The selected cells were transfected with vector encoding pri-miR-122a which was cloned into the tetracycline response element vector, pcDNA 4/myc-His B (Invitrogen), and zeocin resistant colonies were isolated for 2 weeks.
4
Liver Cancer Cell Lines and Reagents
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Human liver cancer cell lines including PLC/PRF/5, QGY7703, Huh7, etc., normal liver cell line Chang and mouse liver cancer cell line Hepa1-6 were all purchased from Cell Bank of the Typical Culture Preservation Committee, Chinese Academy of Sciences (Shanghai, China). PLC/PRF/5, Huh7,Hepa1-6, HepG2, SK-Hep1 and HCCLM3 cells were cultured in the DMEM medium (Invitrogen, Shanghai, China).And Chang, QGY7703, BEL7402 were cultured in RPMI1640 medium (Invitrogen, Shanghai, China). All mediums were supplemented with 10% FBS and 100U/mL penicillin–streptomycin. The following antibodies were used for Western blotting: WIP1 (sc-376257) to detect human samples from Santa Cruz (Shanghai, China); WIP1 (A6204) to detect mouse samples from ABcolonal (Wuhan, China); cleaved PARP1(#9541), cleaved-caspase3 (#9661) and beta-Actin (#4970) from Cell Signaling Technology (Shanghai, China); γH2AX (phospho-Histone H2AX (s139)) (ab81299) from Abcam (Shanghai, China); ki67 (ER1802-31) from Huabio (Hangzhou, China). WIP1 plasmid was kindly provided by Prof. Zhenyu Ju at Hangzhou Normal University. GSK2830371, Diethylnitrosamine (DEN) and TCPOBOP were purchased from Sigma-Aldrich (Shanghai, China). Olaparib (HY-10162) and Veliparib (HY-10129) were purchased from MedChemExpress (Shanghai, China). Other reagents and chemicals don’t list here are commercially available.
5
Culturing Mouse Hepatocyte Cell Lines
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Mouse hepatocellular carcinoma cells (Hepa 1-6) and mouse immortalized hepatocytes (AML12) were purchased from ATCC (Manassas, VA). Hepa 1-6 cells were cultured in Dulbecco’s Modified Eagle’s Medium (DMEM; Thermo Fisher Scientific, Waltham, MA) containing 4mM L-glutamine, 4.5g/L glucose, 1mM sodium pyruvate, and 1.5g/L sodium bicarbonate, supplemented with 10% fetal bovine serum (FBS; Gibco) and 1% penicillin/streptomycin as recommended by ATCC. AML12 cells were cultured in DMEM: F-12 Medium containing 2.5mM L-glutamine, 15mM HEPES, 0.5mM sodium pyruvate, and 1200mg/L sodium bicarbonate, supplemented with 10% FBS, 1% penicillin/streptomycin, 10μg/ml insulin, 5.5μg/ml transferrin, 5ng/ml selenium, and 40ng/ml dexamethasone.
6
Cell Line Authentication and Culture
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The human HCC cell line HCCLM3 and mouse HCC cell line H22 were obtained from the China Center for Type Culture Collection; the human HCC cell line HepG2 and mouse HCC cell line Hepa1-6 were obtained from the Cell Bank of Type Culture Collection Chinese Academy of Sciences. These cell lines were authenticated by short tandem repeat (STR) analysis and tested for mycoplasma contamination. HCCLM3 (abbreviated LM3), HepG2, and Hepa1-6 cells were cultured in DMEM, whereas H22 cells were cultured in RPMI 1640, supplemented with 10% fetal bovine serum (FBS), 2 mM L-glutamine, 100 units/mL penicillin, and 0.1 mg/mL streptomycin (all from Thermo Fisher Scientific, Gibco, Grand Island, NY, USA), and maintained in a humidified incubator with atmosphere containing 5% CO2 at 37°C.
The following reagents were used in this study: fludarabine monophosphate (#F0913; Tokyo Chemical Industry, Tokyo, Japan), MG132 (#M8699; Sigma-Aldrich, St. Louis, MO, USA), human IFN-γ (#Z02915; GenScript, Piscataway, NJ, USA), trypan blue (#ST798; Beyotime Biotechnology, Shanghai, China), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (#M2128; Sigma-Aldrich).
The following reagents were used in this study: fludarabine monophosphate (#F0913; Tokyo Chemical Industry, Tokyo, Japan), MG132 (#M8699; Sigma-Aldrich, St. Louis, MO, USA), human IFN-γ (#Z02915; GenScript, Piscataway, NJ, USA), trypan blue (#ST798; Beyotime Biotechnology, Shanghai, China), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (#M2128; Sigma-Aldrich).
7
Comparative Cell Line Cultivation and Characterization
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The human HCC cell line HCCLM3 and mouse HCC cell line H22 were obtained from the China Center for Type Culture Collection, the mouse HCC cell line Hepa1–6 was obtained from Cell Bank of Type Culture Collection Chinese Academy of Sciences, authenticated by short tandem repeat (STR) analysis, and tested for mycoplasma contamination. HCCLM3 (abbreviated LM3) and Hepa1–6 cells were cultured in high glucose (4.5 g/L) Dulbecco’s modified Eagle’s medium (DMEM) and H22 cells were cultured in Roswell Park Memorial Institute 1640 Medium (RPMI 1640) supplemented with 10% foetal bovine serum, 2 mM l -glutamine, 100 units/mL penicillin, and 0.1 mg/mL streptomycin (all from Thermo Fisher Scientific, Gibco, Grand Island, NY, USA). All cells were maintained in a humidified incubator with an atmosphere containing 5% CO2 at 37 °C.
The following reagents were used: sodium dichloroacetate (#347795; Sigma-Aldrich, St. Louis, MO, USA), trypan blue (#ST798; Beyotime Biotechnology, Shanghai, China), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (#M2128; Sigma-Aldrich).
The following reagents were used: sodium dichloroacetate (#347795; Sigma-Aldrich, St. Louis, MO, USA), trypan blue (#ST798; Beyotime Biotechnology, Shanghai, China), and 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) (#M2128; Sigma-Aldrich).
8
Mouse Hepatocarcinoma Cell Lines Study
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Mouse hepatocarcinoma cell lines Hepa1-6 and H22 were purchased from the China Center for Type Culture Collection (CCTCC, Wuhan, China) and stored at the Three Gorges University Tumor Microenvironment and Immune Therapy Laboratory. The Hepa1-6 cell line was cultured in DMEM (Thermo Fisher, Waltham, MA, USA), and the H22 cell line was cultured in 1640 (Thermo Fisher, Waltham, MA, USA), both supplemented with 10% fetal bovine serum (FBS, Thermo Fisher) and 1% penicillin/streptomycin (Solarbio life sciences, Beijing, China) at 37°C under 5% CO2.
Female BALB/c normal mice (6–8 weeks, 18–20 g) were purchased from the Wuhan Institute of Biological Products Co., Ltd. (Wuhan, China). All mice were kept under specific pathogen-free conditions in the Animal Experimental Center of China Three Gorges University (Hubei, China). All animal experiments were approved by the Animal Experimental Committee of China Three Gorges University (Hubei, China)(ethical approval numbers:2021030CB). All procedures were performed following the guidelines of the Ministry of Science and Technology of the People’s Republic of China.
Female BALB/c normal mice (6–8 weeks, 18–20 g) were purchased from the Wuhan Institute of Biological Products Co., Ltd. (Wuhan, China). All mice were kept under specific pathogen-free conditions in the Animal Experimental Center of China Three Gorges University (Hubei, China). All animal experiments were approved by the Animal Experimental Committee of China Three Gorges University (Hubei, China)(ethical approval numbers:2021030CB). All procedures were performed following the guidelines of the Ministry of Science and Technology of the People’s Republic of China.
9
Culturing Human and Mouse HCC Cell Lines
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The human HCC cell line Hep3B and mouse HCC cell line Hepa 1–6 (syngeneic to C57BL/6 mice) were obtained from the American Type Culture Collection (Manassas, VA, USA). Hep3B cells were cultured in DMEM medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated FBS (Thermo Fisher Scientific) and 1% penicillin–streptomycin. Hepa 1–6 cells were cultured in RPMI 1640 medium (Thermo Fisher Scientific) supplemented with 10% heat-inactivated FBS and 1% penicillin–streptomycin. All cells were incubated at 37°C in a humidified chamber containing 5% CO2.
10
Culturing Murine and Human Cell Lines
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Murine HCC cell lines, including H22 [cultured in RMPI-1640 containing 10% FBS and 1% penicillin/streptomycin mixture (cat. no. 15140-122; Gibco; Thermo Fisher Scientific, Inc.)] and Hepa1–6 (cat. no. CL-0105; cultured DMEM containing 10% FBS and 1% penicillin/streptomycin mixture), were obtained from the American Type Culture Collection. Murine peripheral macrophages (PMs) and bone marrow-derived macrophages (BMDMs) were cultured in DMEM containing 10% FBS and 1% penicillin/streptomycin mixture and placed at 37°C in a humidified incubator containing 5% CO2. The human monocyte cell line THP-1 (cat. no. BNCC100407; BeNa Culture Collection; Beijing Beina Chunglian Institute of Biotechnology) was cultured in RMPI-1640 supplemented with 10% FBS and 1% penicillin/streptomycin mixture. DMEM (cat. no. C11995500CP) was obtained from Gibco; Thermo Fisher Scientific, Inc., RMPI-1640 (cat. no. SH30809.01) was purchased from HyClone; Cytiva and FBS (cat. no. WXBD5226V) was obtained from Sigma-Aldrich; Merck KGaA.
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