The human gastric normal epithelium cell line GES-1 (Cell Bank of the Chinese Academy of Science, Xiangya, China) was cultured in DMEM (Gibco, Grand Island, NY, United States) supplemented with 10% fetal bovine serum (FBS; Gibco), streptomycin (100 g/mL), and penicillin (100 U/mL) at 37 °C in an atmosphere containing 5% CO2. And GC cell lines, including AGS, MKN45, SGC7901, KATOIII, and NCI-N87 (Cell Bank of the Chinese Academy of Science, Shanghai, China), were cultured in DMEM or PRIM-1640 supplemented with 10% FBS, streptomycin, and penicillin. The authenticity of cell lines used in this study had been verified by short tandem repeat profiling.
Fetal bovine serum (fbs)
Manufactured by National Collection of Authenticated Cell Cultures
Sourced in China
Fetal bovine serum (FBS) is a common cell culture medium supplement derived from the blood of bovine fetuses. It provides a complex mixture of proteins, growth factors, and other components that support the growth and proliferation of a wide variety of cell types in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
Rpmi 1640 medium, by National Collection of Authenticated Cell Cultures (3 mentions)
Goat anti mouse igg hrp abs20001, by Absin (3 mentions)
Goat anti rabbit igg hrp abs20002, by Absin (3 mentions)
Penicillin, by National Collection of Authenticated Cell Cultures (2 mentions)
Streptomycin, by National Collection of Authenticated Cell Cultures (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Cleaved caspase 3, by Cell Signaling Technology (2 mentions)
Recombinant human trail, by Merck Group (2 mentions)
Phospho p38, by Cell Signaling Technology (2 mentions)
Caspase 8, by Cell Signaling Technology (2 mentions)
U0126, by AbMole (2 mentions)
Phospho erk, by Cell Signaling Technology (2 mentions)
Caspase 3, by Cell Signaling Technology (2 mentions)
Phospho jnk, by Cell Signaling Technology (2 mentions)
Caspase 9, by Cell Signaling Technology (2 mentions)
Mcl 1, by Cell Signaling Technology (2 mentions)
Bcl 2, by Cell Signaling Technology (2 mentions)
Cleaved caspase 9, by Cell Signaling Technology (2 mentions)
β actin, by Cell Signaling Technology (2 mentions)
15 protocols using fetal bovine serum (fbs)
1
Cell Culture Protocols for Gastric Cell Lines
2
Lentiviral Vector Construction for NIS Expression
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Lv-EF1α-OCT4-IRES-EGFP was kindly provided by the Institute of Molecular Biology, Chinese Academy of Sciences; pcDNA3.1-NIS was obtained from our own library [11 (link)]. The NIS gene was amplified from pcDNA3.1-NIS by PCR using the primers: forward (5’-GCGCGGATCCCGGGTATCGATGGAGGCCGTG-3’) and reverse (5’-CGCGTCTAGATCAGAGGTTTGTAGGTAGTGAGC-3’), digested with XbaI and BamHI, and cloned into the XbaI and BamHI sites of Lv-EF1α-OCT4-IRES-EGFP generating a functional vector featuring NIS under the control of the human elongation factor-1α (EF1α) promoter (the OCT4 transgene of Lv-EF1α-OCT4-IRES-EGFP was replaced with NIS).
HEK293T cell line (Cell Bank of the Chinese Academy of Science, Shanghai, China) was cultured in RPMI-1640 medium supplemented with 10% FBS(Fetal Bovine Serum) and 1% penicillin/streptomycin.
Virus particles were generated by cotransfection of HEK293T cells with Lv-EF1α-NIS-IRES-EGFP and the three packaging plasmids pRsv-REV, pMDIg-pRRE and pMD2G(Biovector Science Lab, Beijing, China). The virus particles were harvested by collecting the cell culture medium at 48 h post-transfection; the supernatants were filtered through 0.45 µm filters, centrifuged at 10,000 g for 15 min and the resulting pellet was resuspended in 100 μl culture medium.
HEK293T cell line (Cell Bank of the Chinese Academy of Science, Shanghai, China) was cultured in RPMI-1640 medium supplemented with 10% FBS(Fetal Bovine Serum) and 1% penicillin/streptomycin.
Virus particles were generated by cotransfection of HEK293T cells with Lv-EF1α-NIS-IRES-EGFP and the three packaging plasmids pRsv-REV, pMDIg-pRRE and pMD2G(Biovector Science Lab, Beijing, China). The virus particles were harvested by collecting the cell culture medium at 48 h post-transfection; the supernatants were filtered through 0.45 µm filters, centrifuged at 10,000 g for 15 min and the resulting pellet was resuspended in 100 μl culture medium.
3
Assessing Cytotoxicity in HEK293 and EJ Cells
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HEK293 cells (DMEM medium supplemented with 10% FBS, commercially purchased from the China Center for Type Culture Collection) and EJ cells were cultured in 96-well cell plates (1×104 cells/well) and treated with 10 μL GVs (OD500 = 2) for 24 hours. Then, CCK-8 solution (Cell Counting Kit-8, Beyotime, Shanghai) was added to the plate and incubated for 1 hour. The absorbance was measured at 450 nm using a microplate reader (SpectraMax M5, Molecular Devices).
4
Hepatocarcinoma Cell Line Cultivation
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Fetal bovine serum (FBS), Dulbecco’s modified Eagle medium (DMEM) and penicillin/streptomycin were purchased from Gibco (GrandIsland, State of New York, USA).The hepatocarcinoma cell lines HepG2, Huh7, HepG2.2.15 cells (derived from HepG2 cells carrying HBV genome) and HepAD38 (replicates HBV under conditions regulated with tetracycline) were obtained from China Center for Type Culture Collection (CCTCC) and grown in DMEM supplemented with 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin. The cells were seeded in 24-well (seeding densities: 2.5 × 105 cells per well), 12-well (seeding densities: 5 × 105 cells per well), 6-well (seeding densities: 1.2 × 106 cells per well), and 6 cm vessels (seeding densities: 2.6 × 106 cells per dish), and were transfected by Lipofectamine 2000 transfection regent following the manufacturer’s instructions.
5
Borax Treatment on HepG2 Cells
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HepG2 cells were obtained from the China Center for Type Culture Collection (Wuhan University, Wuhan, China) and seeded in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum (FBS; cat. no. 10099-141; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) 1 day prior to borax (4 mM; Tianjin Bodi Chemical Co. Ltd., Tianjin, China) treatment in a humidified 5% CO2 incubator at 37°C for either 2 or 24 h. Following 2- or 24-h treatment with 4 mM borax, the culture medium was replenished with fresh media without borax.
6
Colon Cancer Cells and HUVEC Exosome Assays
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HCT-15 (human colon cancer cells) and HCT-15/FU cells were bought from the cell bank of Chinese Academy of Sciences (Shanghai, China) and were cultured in RPMI 1640 medium containing 10% fetal bovine serum (FBS, Gibco, USA). HCT-15/FU cells were supplemented with 3.2 μg/ml 5-FU. Conditioned medium (CM) was collected after cells were cultured in exosome-free FBS medium for 48 h. Human umbilical vein endothelial cells (HUVECs) were purchased from the cell bank of Chinese Academy of Sciences (Shanghai, China) and were cultured in endothelial cell medium (ECM) obtained from ScienCell Research Laboratories. Before treatment of exosomes, cells were cultured in basal medium. All cells were incubated at 37°C with 5% CO2.
7
HL-60 Cell Culture and P-Selectin Binding
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Human promyelocytic leukemia HL-60 cells (Cell Bank of Chinese Academy of Sciences, Shanghai, China), constitutively expressed PSGL-1 as a ligand for P-selectin, were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS), 10 mg/mL streptomycin and 100 units/mL penicillin at 37 °C in a humidified atmosphere of 5% CO2 in air. RPMI-1640 medium, FBS and BSA were purchased from Sigma Chemical Co. (St Louis, MO, USA). streptomycin, penicillin and phosphate buffer saline (PBS) were obtained from Gibco BRL (Grand Island, NY). Recombinant Human P-Selectin/CD62P Fc Chimera Protein (R&D Systems, Minneapolis, MN) is a disulfide-linked homodimer, containing the Fc moiety of human IgG and the extracellular domain of human P-selectin.
8
TRAIL and 5-FU Combination Therapy
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The GES-1, AGS and HGC27 cell lines were purchased from the cell bank of the Chinese Academy of Sciences and cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS). The incubator conditions were 37°C and 5% CO 2 (12) . FBS and RPMI-1640 medium were purchased from Gibco (New York, USA). Recombinant human TRAIL (#T9701) and 5-FU (#F6627) were purchased from Sigma-Aldrich (Munich, Germany). The small-molecule inhibitors U0126 (#M1977), SP600125 (#M2076) and SB202190 (#M2062) were obtained from AbMole (Houston, USA). Primary antibodies against the following proteins were purchased from Cell Signaling Technology (Beverly, MA, USA): TRAIL (#3219), Caspase-3 (#9662), Caspase-8 (#9746), Caspase-9 (#9502), Cleaved Caspase-3 (#9661), Cleaved Caspase-8 (#9496), Cleaved Caspase-9 (#9505), PARP (#9532), Bid (#2002), DR4 (#42533), DR5 (#8047), c-IAP1 (#7065), c-IAP2 (#3130), Bcl-2 (#4223), Mcl-1 (#94296), Erk (#4695), JNK (#9252), p38 (#8690), phospho-Erk (#4370), phospho-JNK (#4668), phospho-p38 (#4511) and β-actin (#4970). The secondary antibodies used in this study included goat anti-mouse IgG-HRP (abs20001) and goat anti-rabbit IgG-HRP (abs20002), both of which were obtained from Absin (Shanghai, China).
9
Culturing RBE Cells in DMEM
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RBE cells (a human CCA cell line) were purchased from the cell bank of the Chinese Academy of Sciences (Shanghai, People’s Republic of China) and cultured with Dulbecco’s Modified Eagle’s Medium containing 10% fetal bovine serum and 100 U/mL penicillin/streptomycin at 37°C in humidified 5% CO2. The above three reagents were purchased from Gibco (New York, NY, USA).
10
TRAIL and 5-FU Combination Therapy
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The GES-1, AGS and HGC27 cell lines were purchased from the cell bank of the Chinese Academy of Sciences and cultured in RPMI-1640 medium containing 10% fetal bovine serum (FBS). The incubator conditions were 37°C and 5% CO 2 (12) . FBS and RPMI-1640 medium were purchased from Gibco (New York, USA). Recombinant human TRAIL (#T9701) and 5-FU (#F6627) were purchased from Sigma-Aldrich (Munich, Germany). The small-molecule inhibitors U0126 (#M1977), SP600125 (#M2076) and SB202190 (#M2062) were obtained from AbMole (Houston, USA). Primary antibodies against the following proteins were purchased from Cell Signaling Technology (Beverly, MA, USA): TRAIL (#3219), Caspase-3 (#9662), Caspase-8 (#9746), Caspase-9 (#9502), Cleaved Caspase-3 (#9661), Cleaved Caspase-8 (#9496), Cleaved Caspase-9 (#9505), PARP (#9532), Bid (#2002), DR4 (#42533), DR5 (#8047), c-IAP1 (#7065), c-IAP2 (#3130), Bcl-2 (#4223), Mcl-1 (#94296), Erk (#4695), JNK (#9252), p38 (#8690), phospho-Erk (#4370), phospho-JNK (#4668), phospho-p38 (#4511) and β-actin (#4970). The secondary antibodies used in this study included goat anti-mouse IgG-HRP (abs20001) and goat anti-rabbit IgG-HRP (abs20002), both of which were obtained from Absin (Shanghai, China).
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