The human gastric cancer lines AGS (moderately-differentiated), MKN-45 (poorly-differentiated), SGC-7901 (moderately-differentiated), MKN-74 (well-differentiated) and MGC-803 (poorly-differentiated) were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). MGC-803 and AGS cells were maintained in Dulbecco's modified Eagle's medium (DMEM; Hyclone, Logan, Utah, USA) supplemented with 10% FBS and the other cell lines were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Hyclone) containing 10% FBS. All cells were maintained at 37°C in a humidified atmosphere containing 5% CO2. Before experiments, gastric cancer cells in log phase growth in six-well plates were cultured in media containing only 1% FBS for 24 h.
Mkn74
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
The MKN74 is a specialized laboratory equipment designed for the cultivation and maintenance of cell cultures. It provides a controlled environment for the growth and propagation of various types of cells. The core function of the MKN74 is to ensure the optimal conditions for cell culture, including temperature, humidity, and gas composition regulation.
Automatically generated - may contain errors
Lab products found in correlation
Mkn 45, by National Collection of Authenticated Cell Cultures (8 mentions)
Hgc 27, by National Collection of Authenticated Cell Cultures (8 mentions)
Mgc 803, by National Collection of Authenticated Cell Cultures (7 mentions)
Sgc 7901, by National Collection of Authenticated Cell Cultures (5 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (4 mentions)
Nci n87, by National Collection of Authenticated Cell Cultures (3 mentions)
Nugc 3, by National Collection of Authenticated Cell Cultures (3 mentions)
Ges 1, by National Collection of Authenticated Cell Cultures (3 mentions)
Bgc823, by National Collection of Authenticated Cell Cultures (3 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Kato 3, by National Collection of Authenticated Cell Cultures (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Sartorius (2 mentions)
Fetal bovine serum (fbs), by Sartorius (2 mentions)
Ham s f 12 medium, by Corning (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Ham s f12 medium, by Thermo Fisher Scientific (1 mentions)
Sun 1, by National Collection of Authenticated Cell Cultures (1 mentions)
Si nc, by Thermo Fisher Scientific (1 mentions)
11 protocols using mkn74
1
Culturing Human Gastric Cancer Cells
2
Gastric Cancer Cell Lines and Tissues
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Seven human GC cell lines (NCI‐N87, MKN74, AGS, NUGC‐3, MKN45, MGC803, and HGC‐27) were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All cell lines were maintained in RPMI‐1640 (Gibco BRL, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (FBS) except AGS in Ham's F12 medium (Cellgro, Manassas, VA, USA) and incubated at an atmosphere containing 5% CO2 at 37 °C. Human GC samples and their corresponding nontumorous gastric tissues were collected at the time of surgical resection at The First Affiliated Hospital of Fujian Medical University (Fuzhou, China) from 2008 to 2010. The tissues were immediately frozen in liquid nitrogen and stored at −80 °C freezer or fixed in 10% formalin for paraffin embedding. All samples were collected with patients’ informed consent, and the study was approved by the institutional review board and regulatory authorities of Fujian Medical University. Clinicopathological classification and staging were determined according to American Joint Committee on Cancer seventh edition of GC TNM staging (Wittekind, 2010 ). No patients had received chemotherapy or radiotherapy before surgery.
3
Gastric Cancer Cell Lines and Samples
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Six human GC cell lines (NCI-N87, MKN74, AGS, NUGC3, MGC803, HGC-27) were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All cell lines were maintained in RPMI-1640 supplemented with 10% FBS except AGS in DMEM/Ham’s F12 medium and incubated at an atmosphere containing 5%CO2 at 37 °C. Gastric adenocarcinoma patients (n = 115) were randomly enrolled in between January 2013 and December 2014 at the Department of General Surgery of the Second Affiliated Hospital of Fujian Medical University. All patients were diagnosed pathologically according to the American Joint Committee on Cancer (AJCC) criteria [26 (link)]. No patient had received chemotherapy or radiotherapy before surgery. Tumor samples and the corresponding noncancerous mucosal tissue were collected from all patients immediately after resection and were frozen in liquid nitrogen. Samples were stored in the Center Laboratory of Second Affiliated Hospital of Fujian Medical University for studies.
4
Culturing Human Gastric Cell Lines
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Human gastric cell lines SGC-7901 and MKN74 were obtained from the Type Culture Collection of the Chinese Academy of Sciences (China). Cells were grown in RPMI 1640 (Rosewell Park Memorial Institute) containing with 10% fetal bovine serum and 1% penicillin–streptomycin in a humidified atmosphere of 95% air and 5% CO2 at 37 °C [24 (link)].
5
Establish Gastric Cancer Cell Lines
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A total of seven human GC cell lines (MKN74, AGS, KATOIII, NUGC3, MGC803, MKN45, and HGC27) were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) and maintained in RPMI-1640 medium (Invitrogen, Carlsbad, CA, USA) except AGS in Ham’s F12 medium (Invitrogen). All media were supplemented with 10% (v/v) fetal bovine serum (Invitrogen).
6
Gastric Cancer Cell Line Maintenance
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The GC cell lines, including NCI-N87, KATO III, MKN74, MKN45, HGC-27, SGC-7901, BGC-823, MGC-803, AGS, SUN-1, HS7467 and the immortalized normal gastric epithelial cell GES-1 were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China) between 2010 and 2015. They were stored in liquid nitrogen tanks in the Research Center of the Fourth Hospital of Hebei Medical University. The GC cells were maintained in RPMI-1640 medium (Gibco, USA) supplemented with 10% Fetal bovine serum (FBS; BI, Israel), 1% penicillin, and 1% streptomycin (Invitrogen, USA) and cultured at 37˚C in a 5% CO2 incubator. All the cells used for experiments had been passaged no more than 20 times and cells were monitored for mycoplasma contamination every 6 months.
7
Gastric Cancer Cell Line Studies
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Human GC cell lines, including MKN-45, MKN-74, AGS and HGC-27, and a normal gastric GES-1 cell line were purchased from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). All of these cell lines were cultured in RPMI-1640 medium (Gibco, Thermo Fisher Scientific, Waltham, USA) containing 10% fetal bovine serum (FBS; Thermo Fisher Scientific), 100 U/mL of penicillin-G, and 100 µg/mL of streptomycin. The cells were maintained in a 37°C humidified atmosphere with 5% CO 2 .
Small interfering (si)-MYBL2 (si-MYBL2-1 5′-CCAAGAG-CACACCTGTTAA-3′; si-MYBL2-2 5′-CCAGAAACAT-GCTGCGTTT-3′), and the scramble siRNA (si-NC, 5′-AC-GTGACACGTTCGGAGAATT-3′) as a negative control (NC), were obtained from Shanghai GenePharma Co., Ltd. (Shanghai, China). The si-MYBL2 (50 nM) and si-NC (50 nM) were transfected into HGC-27 cells (5 × 10 5 cells/ well) using Lipofectamine 2000 (Invitrogen, Carlsbad, USA) according to the manufacturer's instructions. In addition, CDC20 transcript cDNA was inserted into the pCDNA3.1 by Lederer Biological Technology (Guangdong, China), and then transfected into HGC-27 cells (20 µg) to achieve CDC20 overexpression (Ov-CDC20). An empty vector without CDC20 sequence was used as the negative control (OV-NC).
Small interfering (si)-MYBL2 (si-MYBL2-1 5′-CCAAGAG-CACACCTGTTAA-3′; si-MYBL2-2 5′-CCAGAAACAT-GCTGCGTTT-3′), and the scramble siRNA (si-NC, 5′-AC-GTGACACGTTCGGAGAATT-3′) as a negative control (NC), were obtained from Shanghai GenePharma Co., Ltd. (Shanghai, China). The si-MYBL2 (50 nM) and si-NC (50 nM) were transfected into HGC-27 cells (5 × 10 5 cells/ well) using Lipofectamine 2000 (Invitrogen, Carlsbad, USA) according to the manufacturer's instructions. In addition, CDC20 transcript cDNA was inserted into the pCDNA3.1 by Lederer Biological Technology (Guangdong, China), and then transfected into HGC-27 cells (20 µg) to achieve CDC20 overexpression (Ov-CDC20). An empty vector without CDC20 sequence was used as the negative control (OV-NC).
8
Cell Culture Conditions for Gastric Cancer
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Human GC cell lines AGS cells were purchased from the American Type Culture Collection (ATCC). HLECs, HEK-293T, BGC-823, HGC-27, MGC-803, SGC-7901, MKN-74, and MKN-45 were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). HEK-293T cells were cultured in DMEM (Biological Industries, Cromwell, CT, United States), AGS cells were cultured in F12K medium (Biological Industries, Cromwell, CT, United States), and the other cells were cultured in RPMI-1640 medium (Biological Industries, Cromwell, CT, United States). All cells were cultured with 10% fetal bovine serum (FBS; Biological Industries, Cromwell, CT, United States), 100 U/ml penicillin (Invitrogen), 100 μg/ml streptomycin (Invitrogen) and incubated in 5% CO2 at 37°C.
9
Gastric Cancer Cell Line Culture
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Human gastric mucosal epithelial cell line GES-1 and five GC cell lines (AGS, MKN74, MGC-803, HGC27 and MKN45) were purchased from the Type Culture Collection of the Chinese Academy of Sciences (China) and maintained in RPMI-1640 medium (BioInd, Beit Haemek, Israel) supplemented with 10% FBS (BioInd, Beit Haemek, Israel) and 1% penicillin/streptomycin (BioInd, Beit Haemek, Israel) in humidified incubator at 37 °C with 5% CO2. Short tandem repeat (STR) analysis was used to authenticate all cell lines before experiments and all mycoplasma test were negative.
10
Gastric Cancer Cell Line Cultivation
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Nine human GC cell lines (SUN-216, BGC-823, AGS, BGC-803, NUGC4, MKN74, MKN45, SGC-7901, and HGC-27) and a non-malignant gastric mucosal epithelial cell line GES-1 were purchased from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). Cells were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA). The human embryonic kidney (HEK) 293FT cell line was obtained from the American Type Culture Collection (Manassas VA, USA), and cultured and stored according to the provider’s instructions. All cell lines were maintained in a humidified cell incubator at 37 °C with an atmosphere of 5% CO2.
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