All the cell lines used in this study were obtained from ATCC. PANC-1 and Panc 04.03 cells were seeded onto 96-well microplates (5 × 103 cells per well) and cultured for 24 h with DMEM medium (Gibco, 11960-051) supplemented with 10% FBS (Biological Industries, 04-001-1) and 1% Pen/Strep (Beyotime, C0222). After treatment with compounds (TH-Z827, MRTX, or BI-2852), cells were incubated for another 24 h. Cell Counting Kit-8 reagents (Beyotime, C0042) were added and the samples were incubated for another 1 h. Cell viability was measured at OD (450 nm) using a multimode microplate reader (PerkinElmer, EnVision). Data were fitted into an inhibitor-response model to get IC50 values.
Pen strep
Manufactured by Beyotime
Sourced in United States, China
Pen/strep is a commonly used antibiotic mixture composed of penicillin and streptomycin. It is a laboratory reagent used to prevent bacterial contamination in cell culture and other biological experiments.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Sartorius (2 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dmem medium, by Thermo Fisher Scientific (1 mentions)
Hos cells, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Wisent (1 mentions)
Hk 2 cell, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Beyotime (1 mentions)
N2 supplement, by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Corning (1 mentions)
6 well ultra low attachment culture plates, by Corning (1 mentions)
B27 w o vitamin a supplement, by Thermo Fisher Scientific (1 mentions)
Glutamax, by Thermo Fisher Scientific (1 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (1 mentions)
Neurobasal, by Thermo Fisher Scientific (1 mentions)
Fine dissection scissors, by Fine Science Tools (1 mentions)
Rbc lysis buffer, by Solarbio (1 mentions)
Dulbecco modified eagle medium (dmem), by Sartorius (1 mentions)
7 protocols using pen strep
1
Cytotoxicity Evaluation of Compounds
2
Glioma and Glioblastoma Cell Lines
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The low grade glioma cell line H4 and glioblastoma cell line U251 were purchased from Cell Resource Center (CAMS/PUMC, Beijing, China). The glioblastoma cell lines U87 and T98G were purchased from ATCC. All of the cell lines were tested and authenticated. These cell lines were maintained at 37℃ in an atmosphere of 5% CO2 in Dulbecco's Modified Eagle Medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA) and 1% pen-strep (Beyotime, China).
3
Culture of HOS Cells
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HOS cells were purchased from ATCC (USA). Dulbecco’s modified Eagle medium (Gibco, USA), supplemented with 10% fetal bovine serum (Wisent, Quebec, Canada) and 100 μg/mL pen/strep (Beyotime Biotech, Shanghai, China) was used to culture HOS cells at 37°C with 5% CO2.
4
Culturing Human Hepatocellular Carcinoma Cell Lines
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The human HCC cell lines MHCC97-H, MHCC97-L, HepG2, PLC/PRF/5, and HCCLM3 were obtained from the cell bank of the Chinese Academy of Sciences (Shanghai, China). All cells were cultured in DMEM supplemented with 10% FBS (Gibco, Thermo Fisher Scientific, USA) and 1% pen/strep (Beyotime Institute of Biotechnology, Shanghai, China). Cells were cultured at 37°C, 5% CO2, in a humified incubator.
5
HK-2 Cell Glucose Metabolism Regulation
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HK-2 cells from the American Type Culture Collection (Rockville, MD, United States) were cultured in Dulbecco’s modified Eagle’s medium (Gibco, Life Technologies Inc., Rockville, MD, United States; containing 5.5 mM glucose) with 10% fetal bovine serum (Beyotime, China) and 1× Pen/strep (Beyotime). HK-2 cells were treated with high glucose (HG, 30 mM) for 48 h in the absence or presence of 10 μM niclosamide (STAT3 inhibitor) or 10 μM MG132 (proteasome inhibitor). For osmotic control, 5.5 mM glucose and 24.5 mM mannitol (NG) were used.
6
Establishment and Characterization of Lung Cancer PDOs
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PDOs were established and cultured as previously described (81 (link)). Briefly, lung cancer tissues were minced into approximately 0.5–1 mm diameter pieces using fine dissection scissors (Fine Science Tools). Tumor pieces were incubated with RBC lysis buffer (Solarbio, R1010) under gentle rotation for 5 minutes at room temperature to lyse contaminating RBCs. Then, tumor pieces were distributed in ultra-low attachment 6-well culture plates (Corning) with 4 mL per well of PDO medium containing 50% DMEM/F12 (Corning, 10-092-cv), 50% Neurobasal (Thermo Fisher Scientific, 21103049), 1× GlutaMax (Thermo Fisher Scientific, 35050061), 1× NEAAs (Thermo Fisher Scientific, 11140050), 1× PenStrep (Beyotime, C0222), 1× N2 supplement (Thermo Fisher Scientific, 17502048), 1× B27 w/o vitamin A supplement (Thermo Fisher Scientific, 12587010), 1Cat. 2-mercaptoethanol (Thermo Fisher Scientific, 21985023), and 2.5 mg/mL human insulin (Beyotime, P3376-100IU) and placed on an orbital shaker rotating at 120 rpm within a 37°C, 5% CO2, 90% humidity sterile incubator. PDOs were accessed by immunostainings of PanCK and CD31, together with H&E staining.
7
Modulating ITGA2 in Esophageal Cancer
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Human ESCC cell lines KYSE-510, KYSE-150, ECA-109, TE-1, TE-10 and the normal human esophageal epithelial cell line, Het-1A were commercially obtained from Wuhan Fine Biotech Co., Ltd (Wuhan, China) and grown in DMEM (Biological Industries, Israel), containing 100U/mL pen/strep (Beyotime, China) and 10% fetal bovine serum (FBS) (Biological Industries, Israel) in a humidified incubator at 37°C, 5% CO2. ITGA2 was knocked down in the cell lines TE-1 and TE-10, which express high ITGA2 levels. ITGA2 was overexpressed in ECA-109, which has low ITGA2 levels. Puromycin-resistant lentivirus-sh-ITGA2 and lentivirus-ITGA2 were purchased from Genechem (Genechem, China). Virus transfection was carried out according to the manufacturer instructions.
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