Frozen CRC tissues and matched pericarcinomatous tissues were obtained from 18 patients who underwent a CRC resection at Northern Jiangsu People’s Hospital during 2016–2017. Before collecting and using the specimens, all patients provided informed consent and the experiment conformed to the World Medical Association Declaration of Helsinki. Additionally, ethical approval was obtained from the Medical Ethics Committee of Northern Jiangsu People’s Hospital. The specimens were immediately immersed in liquid nitrogen once obtained from patients. The human CRC cell lines HCT116, SW480, and HEK293 were purchased from the Shanghai Institute for Biological Sciences, Chinese Academy of Science. The human colon epithelial cell line (FHC) was purchased from American Type Culture Collection. HCT116 and HEK293 cells were cultured in DMEM (Corning, U.S.A.) with 1% glutamine. While SW480 was cultivated in L15 (Corning, U.S.A.) and FHC in DMEM/F12 (Corning, U.S.A.). All culture media contained 10% fetal bovine serum (Ever Green, China) and 1% penicillin/streptomycin (Invitrogen, U.S.A.). The cells were incubated in 5% CO2 and the temperature maintained at 37°C
Hek293
Manufactured by Corning
Sourced in United States
HEK293 is a specific cell line derived from human embryonic kidney cells. It is commonly used in cell culture and research applications due to its ability to efficiently express recombinant proteins. The HEK293 cell line provides a reliable and well-established system for various experimental purposes.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Corning (6 mentions)
Hek293, by American Type Culture Collection (4 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Fetal bovine serum (fbs), by Corning (3 mentions)
L glutamine, by Corning (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Hct116, by Corning (1 mentions)
Sw480, by Corning (1 mentions)
Dmem f12, by Corning (1 mentions)
Rpmi 1640 media, by Thermo Fisher Scientific (1 mentions)
T75 flask, by Corning (1 mentions)
Dmem media, by Corning (1 mentions)
Rpmi medium, by Corning (1 mentions)
Penicillin, by Merck Group (1 mentions)
Streptomycin, by Merck Group (1 mentions)
L glutamine, by Merck Group (1 mentions)
Tissue culture treated plate, by Corning (1 mentions)
Envision plate reader, by PerkinElmer (1 mentions)
Celltiter glo, by Promega (1 mentions)
13 protocols using hek293
1
Colorectal Cancer Tissue Sampling and Cell Culture
2
Stable Expression of HA-tagged Pol η
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Full-length human Pol η (NCBI NM_006502) complementary DNA was amplified from HEK293 cells by RT-PCR using Coding DNA Sequence (CDS)-specific oligos and cloned in a modified pCDNA3 vector using BamHI and XhoI restriction sites with an in-frame C-terminal HA tag. This vector was then used for generating a stable HEK293 cell line (Pol η-HA HEK293) resistant to G418 (Corning Cat# 30-234-CR) following standard procedure (21 (link)). Stable cells were selected on 400 μg/ml G418 and maintained at a drug concentration of 50 μg/ml. Ectopic expression of Pol η was confirmed by Western blotting of whole cell extract using anti-HA Ab (Cell Signaling Technology Cat# 3724S). All oligos used for cloning are listed in Table S1 .
3
Expanding HEK 293 and INS-832/13 Cells
4
Cell Culture Protocols for Diverse Cell Lines
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HEK293, A-549, H226, and H460 cells were obtained from the American Type Culture Collection (ATCC, USA) and cultured in DMEM (HEK293) or RPMI medium (Corning) supplemented with 10% fetal bovine serum (Corning), 2 mM l -glutamine (Sigma-Aldrich), and antibiotics (penicillin 62.6 µg/ml and streptomycin 40 µg/ml, Sigma-Aldrich). The cells were incubated in a humidified 5% CO2 atmosphere at 37 °C and routinely screened for Mycoplasma contamination.
5
Culturing HEK-293 and HeLa Cells
6
Cell Viability Assay in BJ and HEK293 Cells
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The human cell lines BJ and HEK293 (purchased from the American Type Culture Collection, ATCC, Manassas, VA, USA) were cultured according to ATCC recommendations. BJ is a stable cell line of normal human foreskin fibroblasts; HEK293 is a human embryonic kidney cell line. Approximately 1000 BJ and 400 HEK293 exponentially growing cells were plated per well (in 30 μL) in white polystyrene, sterile flat-bottom 384-well tissue culture–treated plates (Corning, Tewksbury, MA, USA) and incubated overnight at 37 °C in a humidified 5 % CO2 incubator. Compound or fraction stock solutions (in DMSO) were pin-transferred (V&P Scientific, San Diego, CA, USA) the following day. Plates were returned to the incubator for 72 h and equilibrated at room temperature for 20 min before addition of 25 μL Cell Titer Glo (Promega) to each well. Plates were shaken on an orbital shaker for 2 min at 500 rpm. Luminescence was read on an Envision plate reader (Perkin Elmer, Waltham, MA, USA) after 15 min. EC50 values were calculated with RISE by using a four-parameter logistic equation.
7
Cell Line Characterization and Maintenance
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HCC827, PC9, A549, H1299, LO2, and HEK293 cells were obtained from ATCC (American Type Culture Collection). NCI-H1975 and MRC5 cells were purchase from the cell bank of the Chinese Academy of Sciences (Shanghai, China). NCI-H1975, HCC827, and PC9 cells were maintained in RPMI 1640 (Gibco, USA) and A549, MRC5, LO2, and HEK293 in DMEM (Corning, USA) supplemented with 10% FBS (PAN-Biotech, Germany) and antibiotics (100 U/ml Penicillin-Streptomycin, Gibco) at 37 °C in an atmosphere of 5% CO2 humidified environment. All cell lines are regularly authenticated by checking cell morphology and growth rates. If mycoplasma is contamination free is verified by PCR analysis (TaKaRa, Cat. #6601, Japan).
8
Maintenance of NSCLC and HEK-293 Cell Lines
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H1299 (Passage number #17) A549 (Passage number #23) (NSCLC) and human embryonic kidney 293 (Passage number #12) (HEK-293) cell lines were obtained from ATCC (Manassas, VA, USA). H1299 and A549 cell lines were maintained in RPMI-1640 medium (Corning Inc., Corning, NY, USA) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals, Minneapolis, MN, USA), 5% sodium pyruvate (Corning Inc., Corning, NY, USA), and 5% penicillin-streptomycin (Corning Inc., Corning, NY, USA) at 5% CO2 and 37 °C. HEK-293 cell line was maintained in Dulbecco’s Modified Eagle’s Medium (Corning Inc., Corning, NY, USA) supplemented with 10% heat-inactivated FBS (Atlanta Biologicals, Minneapolis, MN, USA) and 5% penicillin-streptomycin (Corning Inc., Corning, NY, USA) at 5% CO2 and 37 °C. All cells were grown to 85–90% confluency prior to splitting or being used for experiments.
9
Culturing HEK293 Cells
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Human embryonic kidney cells (HEK293) were purchased from ATCC. HEK293 cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, Corning) supplemented with 10% fetal bovine serum (FBS, Corning) and 1% antibiotic-antimycotic solution (Gibco). Cells were passaged approximately three times per week and maintained in a humidified incubator at 37°C with 5% CO2.
10
Culturing HEK-293 and HeLa Cells
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HEK-293 and HeLa cells were purchased from American Type Culture Collection (ATCC; Manassas, VA). We cultured HeLa and HEK-293 cells in Dulbecco’s Modified Eagle’s Media (DMEM; 4.50 g/l glucose, 0.58 g/l L-glutamine, 0.11 g/l sodium pyruvate; Corning, Corning, NY) supplemented with 10% fetal bovine serum (FBS; Thermo Scientific, Waltham, MA), but without antibiotics. The cells were maintained in a humidified 37 °C incubator supplied with 5% CO2.
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