Human laryngeal cell carcinoma line human epithelial type 2 (Hep-2) was obtained from the American Type Culture Collection (ATCC; Rockville, MD, USA) and cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), 100 U/ml penicillin, and 100 μg/ml streptomycin (all from Invitrogen, Grand Island, NY, USA) at 37°C in a 5% CO2 incubator. The cisplatin-resistant Hep-2 (Hep-2/R) cells were developed from the parental Hep-2 and cultured in DMEM supplemented with 2 μM cisplatin (Sigma-Aldrich, St. Louis, MO, USA) at 37°C with 5% CO2. To exclude the inference of residual cisplatin, Hep-2/R cells were cultured in cisplatin-free medium for 2 weeks before experiments.
Hep 2
Manufactured by Merck Group
Sourced in United States
The Hep-2 is a type of laboratory equipment used for diagnostic testing. It is designed to detect and identify the presence of specific antigens or antibodies in biological samples. The core function of the Hep-2 is to provide accurate and reliable results for clinical analysis.
Automatically generated - may contain errors
Lab products found in correlation
Hep 2, by American Type Culture Collection (4 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Cisplatin, by Merck Group (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Dmem medium, by Merck Group (1 mentions)
Pgl3 basic vector, by Promega (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Fosinopril, by Merck Group (1 mentions)
Bp151 100, by Thermo Fisher Scientific (1 mentions)
4 protocols using hep 2
1
Establishment and Maintenance of Cisplatin-Resistant Human Laryngeal Cancer Cell Line
2
Cell Culture of Cancer and Kidney Lines
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Human laryngeal cancer cell lines Hep-2 and TU212 and human embryonic kidney cell line HEK293 were obtained from American Type Culture Collection (ATCC). Hep-2 and TU212 cells were grown in DMEM medium (Sigma-Aldrich, St. Louis, MO, USA), supplemented with 10% fetal bovine serum, 1% penicillin and streptomycin (Hyclone, Logan, UT, USA). Cells were cultured in an incubator containing a humidified atmosphere with 5% CO2 at 37 °C.
3
Investigating ACE's Role in Laryngeal Cancer
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The human laryngeal cancer cell line Hep-2 was purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum and 100 μg/ml penicillin/streptomycin at 37°C with 5% CO2.
ACE gene were cloned from human genomic DNA into the pGL3 basic vector (Promega, E1751). The cells were seeded in 96-well plates. When the concentration reached 80~90%, the cells were transfected with plasmids pGL3 containing ACE and empty plasmids pGL3 using Lipofectamine 2000 (Invitrogen 11668027) according to the manufacturer’s instructions. Moreover, in order to reinforce the role of ACE on the progression of laryngeal cancer, some of the Hep-2 cells transfected with empty plasmids pGL3 were treated with Fosinopril (Sigma) at the concentration of 10 μmol/L for 48 h. The experiment was performed in triplicate.
ACE gene were cloned from human genomic DNA into the pGL3 basic vector (Promega, E1751). The cells were seeded in 96-well plates. When the concentration reached 80~90%, the cells were transfected with plasmids pGL3 containing ACE and empty plasmids pGL3 using Lipofectamine 2000 (Invitrogen 11668027) according to the manufacturer’s instructions. Moreover, in order to reinforce the role of ACE on the progression of laryngeal cancer, some of the Hep-2 cells transfected with empty plasmids pGL3 were treated with Fosinopril (Sigma) at the concentration of 10 μmol/L for 48 h. The experiment was performed in triplicate.
4
Yeast Adherence to Epithelial Cells
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Adherence was assessed using Hep-2 (American Type Culture Collection ATCC CCL23) epithelial cells. Yeast cells in exponential phase were added to confluent Hep-2 cells in RPMI 1640 medium without serum (Sigma R1383) at various multiplicities of infection (MOI). Contact between Hep-2 and yeast cells was initiated by brief (1 to 2 min) centrifugation (500 x g)67 (link). After incubation for 1 hour, nonadherent cells were removed by five washes in PBS. Adherent cells were recovered by lysis of the monolayer using 0.5 mL of 0.1% triton (Fisher Scientific BP151-100), 0.5 % SDS (Fisher Scientific BP1311-200), 10 mM EDTA in PBS. Adherent yeast cells were scraped off the well, and serial dilutions were made in distilled water, which were then cultured on YPD agar plates for colony count enumeration. Percentage of adherence was calculated by dividing colony forming units (CFUs) of adherent cells over CFU of input cells and multiplied times 100.
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