The K562 (Chronic myelogenous leukemia) and Hep3B (Hepatocellular carcinoma) tumor cell lines were obtained from ATCC (Manassas, VA, USA) and used as positive tumor controls, as K562 express WT1 and PRAME and Hep3B expresses BIRC5. K562 cells were cultured in Iscove’s Modified Dulbecco’s Medium (ATCC; Cat No. 30-2005) containing 10% FBS in a 5% CO2 incubator at 37°C. Hep3B cells were cultured in Eagle’s Minimum Essential Medium (ATCC; Cat No. 30-2003) containing 10% FBS in a 5% CO2 incubator at 37°C.
Hep3b
Manufactured by American Type Culture Collection
Sourced in United States, China, Japan, United Kingdom, Australia
Hep3B is a human cell line derived from a liver cancer (hepatocellular carcinoma) patient. It is a widely used in vitro model for liver disease research and drug development.
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Lab products found in correlation
Hepg2, by American Type Culture Collection (485 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (196 mentions)
Plc prf 5, by American Type Culture Collection (137 mentions)
Sk hep1, by American Type Culture Collection (134 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (114 mentions)
Smmc 7721, by American Type Culture Collection (73 mentions)
Hep3b, by Thermo Fisher Scientific (64 mentions)
Snu 449, by American Type Culture Collection (63 mentions)
Mhcc97h, by American Type Culture Collection (61 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (59 mentions)
Streptomycin, by Thermo Fisher Scientific (59 mentions)
Penicillin, by Thermo Fisher Scientific (52 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (51 mentions)
Hek293t, by American Type Culture Collection (51 mentions)
Rpmi 1640, by Thermo Fisher Scientific (45 mentions)
Hcclm3, by American Type Culture Collection (42 mentions)
Snu 387, by American Type Culture Collection (41 mentions)
Mhcc97l, by American Type Culture Collection (39 mentions)
Bel 7402, by American Type Culture Collection (36 mentions)
Hepg2, by Thermo Fisher Scientific (36 mentions)
819 protocols using hep3b
1
Culturing K562 and Hep3B Tumor Cell Lines
2
Standardized Hepatocyte Cell Culture
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HepG2, Hep3B and Huh7 hepatocyte-derived cells, and HeLa cells were purchased from the ATCC (Rockville, MD, USA). HepG2 cells and Hep3B (ATCC (Rockville, MD, USA) were cultured in Dulbecco's Modified Eagle Medium (DMEM) with 10% FBS and 50 units/ml penicillin and 50 μg/ml streptomycin, and Huh7 were grown in RPMI 1640 [48 (link)].
3
Culturing Human Liver Cancer Cell Lines
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The HCC cell lines SNU387, SNU398, SNU449, and Hep3B were obtained from the ATCC (Manassas, VA, USA). SNU398 and SNU449 cells were cultured with Roswell Park Memorial Institute 1640 medium with 10% heat-inactivated fetal bovine serum, SNU387 cells were maintained in Roswell Park Memorial Institute 1640 medium with 10% fetal bovine serum, and Hep3B cells were cultured with ATCC-formulated Eagle's minimum essential medium with 10% fetal bovine serum. One hundred units of penicillin and streptomycin were added to each cell culture media.
4
Cell Lines Authentication and Culture
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Human HepG2, HEK293T, CL-48, THLE-2, Hep3B, SNU449, PLC/PRF/5 cell lines were obtained from American Type Culture Collection (ATCC), Huh7 and MHCC97H cell lines were given from Professor Wendong Huang (City of Hope, Duarte, CA). HepG2, HEK293T, Huh7 and MHCC97H were all cultured in DMEM medium (Thermo Fisher Scientific), SNU-449 were all cultured in RPMI-1640 medium (Thermo Fisher Scientific), Hep3B, CL-48 and PLC/PRF/5 were all cultured in EMEM medium (ATCC), THLE-2 were all cultured in BEGM Bullet Kit (Lonza), supplemented with 10% fetal bovine serum (Gemini Bio-Products) and 1% penicillin/streptomycin (Thermo Fisher Scientific) at 37 °C in a 5% CO2 humidified incubator. All cell lines were identified by STR cell authentication and routinely tested for mycoplasma contamination using PCR Mycoplasma Detection Kit (G238, Applied Biological Materials).
5
Cell Line Validation and Authentication
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Cell lines were obtained from the following sources and validated by STR analysis: HepG2 (ATCC HB-8065), FOCUS [35] , Mahlavu[36] , Hep3B (ATCC HB-8064), Hep3B-TR [37] , Huh7 (JCRB JCRB0403), SkHep1 (ATCC HTB- 52), PLC (ATCC CRL-8024), MDA-MB-453 (ATCC HTB-131), HCC1937 (ATCC CRL-2336), BT-20 (ATCC HTB-19), T47D (ATCC HTB-133), CAMA- 1 (ATCC HTB-21), HCT116 (ATCC CCL-247), HT29 (ATCC HTB-38), SW620 (ATCC CCL-227).
6
Culturing Immortalized Liver Cells
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The immortalized normal liver epithelial cells, THLE-3, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and were cultured under the conditions stated by the manufacturer. The HCC cell lines: HepG2, Hep3B, MHCC97H, MHCC97L, BEL-7402, Huh7, SMMC-7721, PLC/PRF/5, and QGY-7703 were kindly donated by Prof. Qian Wang (Sun Yat-Sen University, Guangzhou, China), which HepG2, Hep3B and PLC/PRF/5 were purchased from ATCC, and MHCC97H, MHCC97L, BEL-7402, Huh7, SMMC-7721 and QGY-7703 were purchased from Shanghai Yansheng industrial Co. (Shanghai, China). The HCC cells were grown in Dulbecco’s modified Eagle’s medium (DMEM, Gibco-BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco-BRL) supplemented with 100 U/ml penicillin and 100 μg/ml streptomycin (Gibco-BRL), at 37°C in a 5% CO2 atmosphere in a humidified incubator.
7
Establishment of Luciferase-Labeled Liver Cancer Cell Lines
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SNU-449 and Hep3B cell lines were all purchased from American Type Culture Collection (ATCC, USA) in May 2019. Hep3B [5] was cultured in EMEM (ATCC, USA) with 10% fetal bovine serum (FBS, Gibco, USA) and SNU-449 was cultured in RPMI-1640 medium, with 10% heat-inactivated FBS, respectively. Both of them were cultured in the same condition of humidified incubator at 37°C with 5% CO2. Hep3B-luc and SNU-449-luc were then acquired by CMV-Firefly luciferase-IRES-Puro lentivirus transfection. For details, cell selection was conducted for at least 12 days with 1μg/ mL puromycin. Stable fluorescence signal of both cell lines was confirmed by 96 microplate luminometer (Promega, USA). Two cell lines were both characterized using STR (Short Tandem Repeat) analysis for identity verification of human cell lines in 2019.
8
Establishment of DDP-Resistant HCC Cell Lines
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Human HCC cells (Huh7 and Hep3B) were purchased from the American Type Culture Collection and were incubated with gradually increasing concentrations of DDP for 12 months to obtain DDP-resistant Huh7 (Huh7/DDP) and DDP-resistant Hep3B (Hep3B/DDP) cell lines. The cells were subsequently cultured in RPMI-1640 medium (Invitrogen; Thermo Fisher Scientific, Inc.) supplemented with 10% FBS (Gibco; Thermo Fisher Scientific, Inc.) at 37°C (5% CO2).
9
Hepatocellular Carcinoma Cell Lines and Animal Models
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Human HCC cell lines (Hep G2, SMMC-7721, and Hep 3B) and normal hepatocyte cell line (LO 2) were obtained from American Type Culture Collection (ATCC), cultured at 37 °C under a humidified atmosphere of 5% CO2 in DMEM (Hep G2, SMMC-7721, and LO 2) or 1640 (Hep 3B) medium supplemented with 10% fetal bovine serum, 100 IU/mL penicillin, and 100 μg/mL streptomycin.
Male BALB/c mice (18–22 g), BALB/c nude mice (18–22 g), and Sprague-Dawley (SD) rats (200–250 g) were purchased from Nanjing Qing Longshan Animal Breeding Company (Nanjing, China). The animals were housed with ad libitum access to food and water at 25 °C and relative humidity of 55%. Animal study protocols were approved by Animal Ethics Committee of Nanjing University of Chinese Medicine, and animal studies were performed in accordance with the Guide for Care and Use of Laboratory Animals.
Male BALB/c mice (18–22 g), BALB/c nude mice (18–22 g), and Sprague-Dawley (SD) rats (200–250 g) were purchased from Nanjing Qing Longshan Animal Breeding Company (Nanjing, China). The animals were housed with ad libitum access to food and water at 25 °C and relative humidity of 55%. Animal study protocols were approved by Animal Ethics Committee of Nanjing University of Chinese Medicine, and animal studies were performed in accordance with the Guide for Care and Use of Laboratory Animals.
10
Surgical Treatments of Hepatocellular Carcinoma
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This research comprised 46 HCC patients who received surgical treatments during 2014 and 2017, along with adjacent normal tissues, were obtained and verified by a trained pathologist. These patients were not exposed to any treatments (chemotherapy or radiotherapy) before surgery. Informed consent was taken from all participating patients. Human normal liver cell line THLE‐3 and HCC cell lines HepG2, HepG2.2.15, Hep3B, Huh7, and Hep3B cells were purchased from the American Type Culture Collection. Dulbecco's Modified Eagle's Medium (DMEM, Gibco, United States) supplemented with 10% fetal bovine serum (FBS, Gibco) was procured for cultivating THLE‐3 and all HCC cells. The cells were maintained under the standard conditions (37°C and 5% CO2). Detailed clinicopathological features are described in Table 1 .
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