The trophoblast cell lines HTR-8/SVneo (HTR-8) and JEG3 were grown in DMEM/F-12 medium (Gibco, USA), the human monocyte cell line THP-1 was cultured in PRIM-1640 medium (Gibco), and Raw 264.7 macrophages were grown in DMEM/high glucose medium (Gibco) supplemented with 10% fetal bovine serum (FBS) (Gibco) at 37 °C in 5% CO2. For M0 macrophage polarization, 5 × 105/mL THP-1 cells were cultured in PRIM-1640 with 50 ng/mL phorbol 12-myristate 13-acetate (PMA; Sigma-Aldrich, USA) for 24 h. For M1 macrophages, PMA-stimulated THP-1 cells or Raw 264.7 macrophages were stimulated with 100 ng/mL lipopolysaccharide (LPS; Sigma-Aldrich) and 20 ng/mL IFN-γ (PeproTech, USA), while IL-4 and IL-13 were applied to stimulate for M2 macrophages 16 , 28 (link).
Prim 1640 medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
The PRIM-1640 medium is a cell culture medium designed for the growth and maintenance of various cell lines. It provides a balanced formulation of essential nutrients, vitamins, and growth factors to support the optimal growth and proliferation of cells in vitro.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (7 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Ifn γ, by Thermo Fisher Scientific (1 mentions)
Dmem high glucose medium, by Thermo Fisher Scientific (1 mentions)
Dmem f12 medium, by Thermo Fisher Scientific (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Gsk j1, by Selleck Chemicals (1 mentions)
Epz005687, by Selleck Chemicals (1 mentions)
Dibutyryl camp db camp, by Merck Group (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
24 well plate, by Corning (1 mentions)
Nci n87, by Shanghai Cell Bank (1 mentions)
Lipofectamine 3000, by Thermo Fisher Scientific (1 mentions)
Sgc 7901, by Shanghai Cell Bank (1 mentions)
Ges 1, by Shanghai Cell Bank (1 mentions)
Bgc 823, by Shanghai Cell Bank (1 mentions)
Minimum essential medium (mem), by Merck Group (1 mentions)
10 protocols using prim 1640 medium
1
Macrophage Polarization Protocol
2
ESCC Cell Line Transfection Protocol
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Human ESCC cell lines KYAE‐1, TE8, TE5 and FLO‐1 were purchased from the Chinese Type Culture Collection, Chinese Academy of Sciences (Shanghai, China). All cell lines were cultured in PRIM 1640 medium (Gibco) supplemented with 10% fetal bovine serum (FBS, Gibco) and 100 U/mL penicillin/streptomycin (Sigma) at 37°C in a humidified incubator with 5% CO2 in atmosphere. Cells were transfected with linc01014 mimic, si‐linc01014 or negative control (NC) using lipofectamine 2000 (Invitrogen) according to manufacturer's protocol.
3
Cultivation of KIRC Cell Line 786-O
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Human KIRC cell line 786-O were conserved in our laboratory. Cells were cultured in PRIM 1640 medium (Gibco, Carlsbad, CA, USA). The cell media contained 10% fetal bovine serum (FBS, HyClone, Invitrogen), 100 U/ml penicillin and 100 mg/ml, cells were maintained in a humidified incubator at 37 °C with 5% CO2.
4
Investigating GBM Cell Culturing and Treatments
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DBTRG‐05MG cells were cultured at 37 °C under 5% CO2 in PRIM‐1640 medium (Gibco, Thermofisher Scientific, Rockford, IL, USA) supplemented with 10% FBS (Gibco) and penicillin/streptomycin (HyClone, Thermofisher Scientific, Rockford, IL, USA). The cell line was purchased from American Type Culture Collection (ATCC, Manassas, VA, USA).
Primary patient‐derived GBM cells and glioma stem cells (GSCs) were provided by Professor Z. Chen (Department of Neurosurgery/Neuro‐oncology, Sun Yat‐sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine). Primary GBM cells were cultured in DMEM/F12 (Gibco) supplemented with 10% FBS and penicillin/streptomycin.
The following reagents were used in this study: dibutyryl‐cAMP (dbcAMP; 100 mm , dissolved in double distilled water, D0627‐1G; Sigma‐Aldrich, St. Louis, MO, USA), KT5720 (10 mm , dissolved in DMSO, B9002; APExBIO Chemicals, Houston, TX, USA), GSK J1 (10 mm , dissolved in DMSO, S7581; Selleck Chemicals,Houston, TX), and EPZ005687(10 mm , dissolved in DMSO, S7004; Selleck Chemicals, Houston, TX, USA).
Primary patient‐derived GBM cells and glioma stem cells (GSCs) were provided by Professor Z. Chen (Department of Neurosurgery/Neuro‐oncology, Sun Yat‐sen University Cancer Center, State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine). Primary GBM cells were cultured in DMEM/F12 (Gibco) supplemented with 10% FBS and penicillin/streptomycin.
The following reagents were used in this study: dibutyryl‐cAMP (dbcAMP; 100 m
5
Base Editor Transfection in HEK293T, N2a, and Hct116 Cells
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HEK293T and mouse neuroblastoma N2a cells were cultured in DMEM (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) incubated at 37°C in an atmosphere of 5% CO2. Hct116 cells (human colon cancer cell) were cultured in the PRIM-1640 medium (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Gibco) under the same culture conditions. The cells were seeded onto 24-well plates (Corning) and transfected with 1,500 ng base editors and 500 ng gRNA expression plasmids by EZtrans (Shanghai life iLAB BIO Technology) per well following the manufacturer’s instructions. After transfection for 72 h, the cells were collected and sorted about 10,000 fluorescent-positive cells using flow cytometry for PCR amplification and sequencing.
6
Daam1 Modulation in Gastric Cancer
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Human gastric cancer cell lines (AGS, SGC-7901, NCI-N87, BGC-823) and the human normal gastric mucosa cell line GES-1 were obtained from Shanghai Cell Bank, Chinese Academy of Sciences. All these cell lines were incubated in PRIM-1640 medium supplemented with 10% fetal bovine serum (Gibco, USA).
The Daam1 overexpression plasmid and the corresponding negative control plasmid (pCMV6) were obtained from Origene (Rockville, USA). Transfection was performed using Lipofectamine 3000 (Invitrogen, USA). Daam1 small interfering siRNA was purchased from Dharmacon (siRNA1 target sequence: GAGAUAAGUUUGUGUCUGU; siRNA2 target sequence: GUACGAAUGUUGGUUAAUG) (Horizon, Lafayette, CO, USA). siRNA transfection was performed using Dharmafect1.
The Daam1 overexpression plasmid and the corresponding negative control plasmid (pCMV6) were obtained from Origene (Rockville, USA). Transfection was performed using Lipofectamine 3000 (Invitrogen, USA). Daam1 small interfering siRNA was purchased from Dharmacon (siRNA1 target sequence: GAGAUAAGUUUGUGUCUGU; siRNA2 target sequence: GUACGAAUGUUGGUUAAUG) (Horizon, Lafayette, CO, USA). siRNA transfection was performed using Dharmafect1.
7
Time-Course TGF-β1 Stimulation of hOSCC Cells
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All cell lines were grown at 37°C and 5% CO2. Human HSC-4 SCC cells (JCRB0624) were cultured in Eagle's minimum essential medium (MEM; Sigma-Aldrich) supplemented with 10% fetal bovine serum (FBS; Gibco BRL). SAS cells (JCRB0260) were cultured in PRIM1640 medium (Gibco BRL) supplemented with 10% FBS. HO-1-N1 cells (JCRB0831) were cultured in Dulbecco's modified Eagle's medium (DMEM) and Ham's F-12 medium (1:1; Gibco BRL) with 10% FBS. The culture medium was removed and replaced with serum-free medium 24 h prior to the TGF-β1-stimulated experiments. For time-course experiments, 2.0×105 hOSCC cells were cultured in 500 µl of medium without serum containing 10 ng/ml TGF-β1, for 1 to 48 h in 12 or 24-well tissue culture plates.
8
Immortalized Gastric Cell Lines for IM Model
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GES-1, an immortalized gastric epithelial cell line, was used mainly to establish the IM model which was described previously [26 (link)]. AGS, MKN45 and AZ521 are gastric cancer cell lines. HCT-116 is one kind of colon cancer cell line. All of the cell lines were purchased from ATCC and resuscitated within 6 months and they were also tested negative for mycoplasma contamination. Gastric cells were cultured in PRIM-1640 medium (Gibco, US), while colon cells were cultured in DMEM medium (Gibco, US) and all cell lines were cultured with 10% fetal bovine serum (Biological Industries, Israel), 100 mg/ml streptomycin and 100 U/ml penicillin. Deoxycholic acid (DCA) is a major hydrophobic BA with strong cytotoxicity and it was purchased from BiocytoSci (USA).
9
Salidroside Modulates Apoptosis and Signaling Pathways
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PRIM1640 medium, penicillin‐streptomycin solution and trypsin‐EDTA were purchased from Gibco (California, USA). Fetal bovine serum (FBS) was purchased from Lonsera (Montevideo, Uruguay). Salidroside (pure ≥98%) was purchased from Tauto Biotech Co., Ltd. (Shanghai, China) and its product ID is E‐0069. Salidroside was dissolved in phosphate buffer solution (PBS) and filtered through a 0.22‐μm filter before use. Annexin V apoptosis detection kit was purchased from NeoBioscience (Shenzhen, China). Antibodies were obtained from the following sources: rabbit anti‐MMP9 (#ab38898), anti‐pSTAT3 (Tyr705) (#ab76315), anti‐STAT3 (#ab68153), anti‐JAK2 (#ab108596) and anti‐pJAK2 (Y1007 + Y1008) (#ab32101) from Abcam, rabbit anti‐MMP2 (#AF0577) from Affinity, mouse anti‐Bcl‐2 (C‐2) (#sc‐7382) from Santa Cruz Biotechnology, rabbit anti‐Bax (#GB11007) from Servicebio, rabbit anti‐cleaved caspase 3 (#9664) from Cell Signaling Technology, mouse anti‐β‐Actin (#LocusID60) from OriGene, rabbit anti‐GAPDH (#10494‐1‐AP) and HRP‐conjugated secondary anti‐rabbit IgG antibody (#SA00001‐2) from Proteintech.
10
Bile Acid-Induced Gastric Cell Model
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The BA-induced IM cell model was described previously [18] . GES-1 and AGS (purchased from ATCC) were cultured in PRIM-1640 medium (Gibco, USA) with 10% fetal bovine serum (Gibco, USA), 100 mg/ml streptomycin and 100 U/ml penicillin. The primary cells were incubated in ICell Primary Epithelial Cell Culture System (icellbioscience, China) with 2% fetal bovine serum (Biocytosci, USA). Deoxycholic acid (DCA) was chosen for the following experiments as it is a major hydrophobic bile acid with potent cytotoxicity (BiocytoSci, USA).
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