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Sgc 7901

Manufactured by Merck Group
Sourced in United States

The SGC-7901 is a laboratory equipment product manufactured by the Merck Group. It is designed to perform specific functions within a research or testing environment. The core function of the SGC-7901 is to provide precise and reliable measurements or data collection, but a detailed description of its intended use or performance capabilities cannot be provided in an unbiased and factual manner without the risk of extrapolation.

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13 protocols using sgc 7901

1

Establishment of Cisplatin-Resistant Gastric Cancer Cell Lines

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Human gastric cancer cell lines SGC-7901 and BGC-823 were purchased from the Type Culture Collection of the Chinese Academy of Sciences, Shanghai, China. Both the cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum (FBS), 100U/ml penicillin, and 100μg/mL streptomycin (Life Technologies, Gibco, MD, USA). Cells were grown at 37°C, in a humidified incubator with 5% CO2. Cisplatin resistant gastric cancer cells lines SGC-7901/DDP and BGC-823/DDP were produced from parental SGC-7901 and BGC-823cells by persistence gradient exposure to Cisplatin for about 12 months, which were subjected to increasing concentrations from 0.05μg/mL until the cells acquired resistance to 1μg/mL of Cisplatin (Sigma-Aldrich, St. Louis, MO, USA). Prior to each experiment, SGC-7901/DDP and BGC-823/DDP cells were cultured in original RPMI 1640 medium for 2 weeks.
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2

Establishment of Cisplatin-Resistant Gastric Cancer Cell Lines

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Human gastric cell lines (SGC7901, AGS) were purchased through the Type Culture Collection of the Chinese Academy of Sciences, located in Shanghai, China. The cells were grown in 1640 medium (Gibco, Gaithersburg, MD, USA) containing 10% fetal bovine serum (FBS, Gibco) and incubated at 37° C and 5% CO2. The cisplatin-resistant gastric cancer cell lines SGC-7901/DDP and AGS/DDP were produced from the parental SGC-7901 and AGS cells through persistence gradient exposure to cisplatin for approximately 12 months, and was subjected to increasing concentrations of cisplatin (Sigma-Aldrich, St. Louis, MO, USA) from 0.05μg/mL until the cells acquired resistance to 1μg/mL.
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3

Establishment of DDP-Resistant Gastric Cancer Cell Lines

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BGC823 and SGC7901 human gastric cancer cells were purchased from the Shanghai Cell Collection (Shanghai, China). Cells were cultured in RPMI 1640 medium (Gibco Laboratories, USA) supplemented with 10% (v/v) foetal bovine serum (Gibco Laboratories, USA), 100 μg/mL streptomycin, and 100 U/mL penicillin at 37°C and 5% CO2. DDP was purchased from Sigma (St. Louis, MO, USA).
DDP-resistant SGC7901/DDP and BGC823/DDP cells were induced from SGC7901 and BGC823 cells, respectively, using a concentration gradient method to increase the half maximal inhibitory concentration (IC50) of DDP. At first, SGC7901 and BGC823 cells were treated with the culture medium containing 0.05 μg/mL DDP for 24 h. Then, the culture medium containing DDP was substituted for the fresh culture medium. When the cell density reached 80%, cells were digested and passage. The cells were treated with 0.05 μg/mL DDP time and again until the cells can be stable passage in such concentration of DDP. Subsequently, the cells were treated with higher concentration of DDP in turn, until the final concentration of DDP reached 1 μg/mL. The cells were cultured in the culture medium containing 1 μg/mL DDP to maintain its drug resistance. Before every experiment, both DDP-resistant gastric cell lines were cultured in drug-free RPMI 1640 medium for 2 weeks.
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4

Culturing Human Gastric Cancer Cell Lines

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Human GC cell lines including HGC-27, AGS, SGC-7901, BGC-823, NCI-N87 and human normal gastric mucosal cells GSE-1 were purchased from Chinese Academy of Sciences Affiliated Cell Resource Center of Shanghai Institute of Life Sciences (Shanghai, China). HGC-27, BGC-823, SGC-7901 and GSE-1 cells were cultured in 90% RPMI 1640 medium (Sigma-Aldrich, USA) supplemented with 10% FBS (Gibco, USA); AGS cells were cultured in 90% F12K medium (Sigma-Aldrich, USA) supplemented with 10% FBS (Gibco, USA); NCI-N87 cells were cultured in 88% RPMI 1640 medium (Sigma-Aldrich, USA) supplemented with 10% FBS (Gibco, USA), 1% glutamax (Invitrogen, USA), and 1% sodium pyruvate (Invitrogen, USA). All these cell lines were incubated in a humidified incubator under 95% air and 5% CO2 condition at 37°C.
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5

Establishing Gastric Cancer Drug Resistance

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The human normal gastric epithelial cells (RGM-1 and GES-1) and GC cell lines (MGC-803, BGC-7901, MKN-45 and HS-746T) utilized in the present study were purchased from the Institute of Cell Research, Chinese Academy of Sciences, Shanghai, China. Short tandem repeat analysis was conducted for the verification of cell lines' authenticity. Cells were cultured in Roswell Park Memorial Institute 1640 medium (RPMI-1640; Gibco, BRL, Carlsbad, CA, USA) with the supplementation of 100 U mL−1 penicillin, 10% fetal bovine serum and 100 mg mL−1 streptomycin (all provided by Sigma-Aldrich, St. Louis, MO, USA), and were incubated in a 37 °C incubator with a humidified atmosphere of 5% CO2.
For the drug resistance experiments, we referred a predecessor's research to conduct a habitual stepwise method to generate cisplatin (DDP) and 5-fluorouracil (5-Fu) resistant SGC-7901 cell line.41 DDP and 5-Fu reagents (Sigma-Aldrich, St. Louis, MO, USA) were prepared for the incubation of cultured cells those treated and non-treated with siRNA or pcDNA vector.
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6

Establishment of Cisplatin-Resistant Gastric Cancer Cell Lines

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The human gastric cancer cell lines SGC7901 and BGC-823 were obtained from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The cells were cultured in RPMI1640 medium (Gibco, Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (FBS, Gibco, Thermo Fisher Scientifics, Inc.) and incubated at 37°C in a humidified incubator with 5% CO2. cisplatin-resistant SGC7901R and BGC823R cells were established by continuous exposure to stepwise-increasing concentrations of cisplatin (Sigma-Aldrich, Merck KGaA, Darmstadt, Germany). Cells that were viable in the cell culture medium with a high concentration of cisplatin (1,000 μg/L) were designated as cisplatin-resistant cells. Parental cells, denoted as SGC7901S and BGC-823S, were cultured under the same conditions without treatment.
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7

Cultivation and Hypoxic Exposure of Gastric Cell Lines

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Human gastric mucosa cell lines GEF-1, HFE-145, human gastric cancer cell lines MKN45, SGC7901, and human embryonic kidney cell line HEK-293T were obtained from the Shanghai Institutes for Biological Sciences Cell Resource Center. HFE-145, MKN45 cells, and HEK-293T cells were maintained in DMEM (Thermo Fisher Scientific Inc, MA, USA) containing 10% fetal bovine serum (FBS, Life Technologies Corporation, Paisley, UK). SGC7901 and GES-1 cells were cultured in RPMI-1640 (Sigma-Aldrich, St Louis, MO, USA) containing 10% FBS. Cells were incubated in a humidified incubator at 37°C with 5% CO2 in an atmosphere of either normoxia (21% oxygen), or hypoxia (1% O2, 5% CO2, and 94% N2 gas mixture) for 24 h.
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8

Establishing 5-FU Resistant Gastric Cancer Cell Line

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Cells of the human gastric cancer cell line SGC-7901 (American Type Culture Collection, Manassas, VA, USA) were cultured in RPMI-1640 medium (Gibco-BRL, Invitrogen Life Technologies, Carlsbad, CA, USA), which was supplemented with 10% heat-inactivated fetal bovine serum, 100 IU penicillin/ml and 100 µg/ml streptomycin (Invitrogen Life Technologies). All of the cells were maintained in a humidified 5% (v/v) atmosphere of CO2 at 37°C. The 5-FU-resistant variant SGC7901/5-FU was obtained from the parent cell line (SGC-7901) by step by step exposure to increasing concentrations of 5-FU (5, 10, 20 and 40 mg/ml in three-day intervals; Sigma-Aldrich, St. Louis, MO, USA). To maintain the 5-FU-resistant phenotype, 5-FU was added to the culture media with a final drug concentration of 1 mg/ml 5-FU for SGC7901/5-FU cells. Lipofectamine 2000 reagent (Invitrogen Life Technologies) was used for the cell transfection in accordance with the manufacturer's instructions.
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9

Cell Culture Conditions for Gastric and Kidney Cell Lines

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Gastric cancer cells MKN45 and SGC7901, human embryonic kidney cell line HEK-293T and human gastric mucosa cell line GES-1 were obtained from Shanghai Institutes for Biological Sciences Cell Resource Center. MKN45 and HEK-293T cells were hatched in high-glucose DMEM (Dulbecco’s modified Eagle’s medium) supplemented with 10% FBS (fetal bovine serum, Life Technologies Corporation, Paisley, UK). SGC7901 and GES-1 cells were maintained in RPMI-1640 (Sigma-Aldrich, St. Louis, MO, USA) supplemented with 10% FBS. All cells were incubated in a humidified incubator at 37°C with 5% CO2. The medium was refreshed every 48 h.
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10

Gastric and Kidney Cell Cultures

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Gastric cancer cell lines MKN45 and SGC7901, human embryonic kidney cell line HEK-293T and human gastric mucosa cell line GES-1 were purchased from the Shanghai Institutes for Biological Sciences Cell Resource Center, Shanghai, China. MKN45 cells and HEK-293T cells were maintained in Dulbecco's modified Eagle's medium containing 10% fetal bovine serum (Life Technologies Corporation, Paisley, UK). SGC7901 and GES-1 cells were cultured in RPMI-1640 (Sigma-Aldrich, St Louis, MO, USA) containing 10% fetal bovine serum. All cells were incubated in a humidified incubator at 37 °C with 5% CO2.
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