All chemicals and enzymes, unless otherwise specified, were purchased from Sigma-Aldrich (St. Louis, MO) and New England Biolabs (Ipswich, WA). Erythro-9-(2-hydroxy-3-nonyl) adenine (EHNA) hydrochloride was purchased from Tocris Bioscience (Ellisville, MO). 15N3-cytidine-5′-triphosphate was obtained from Sigma-Aldrich (St. Louis, MO) and all other stable isotope-labeled nucleoside starting materials were from Cambridge Isotope Laboratories (Tewksbury, MA). Mouse tissues were obtained from 19-21 week old animals. The HEK293T embryonic kidney cells, MCF7 human breast cancer cells,HCT116 human colon cancer cells, HeLa human cervical cancer cells, WM-266-4 human melanoma cells, and cell culture reagents were purchased from ATCC (Manassas, VA).
Hek293t embryonic kidney cells
Manufactured by American Type Culture Collection
Sourced in United States
HEK293T embryonic kidney cells are a widely used cell line derived from human embryonic kidney cells. These cells are known for their ability to efficiently express recombinant proteins and are commonly used in various research and biotechnology applications.
Automatically generated - may contain errors
Lab products found in correlation
143b osteosarcoma cells, by American Type Culture Collection (3 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Erythro 9 2 hydroxy 3 nonyl adenine ehna hydrochloride, by Bio-Techne (1 mentions)
Mcf 7 human breast cancer cells, by American Type Culture Collection (1 mentions)
Hct116 human colon cancer cells, by American Type Culture Collection (1 mentions)
Hela human cervical cancer cell, by American Type Culture Collection (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Avantor (1 mentions)
Penicillin streptomycin solution, by Corning (1 mentions)
H1792, by American Type Culture Collection (1 mentions)
H1975, by American Type Culture Collection (1 mentions)
H2228, by American Type Culture Collection (1 mentions)
Hcc 366, by American Type Culture Collection (1 mentions)
H1993, by American Type Culture Collection (1 mentions)
H1299, by American Type Culture Collection (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Ccl 171, by American Type Culture Collection (1 mentions)
Mrc 5, by American Type Culture Collection (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
U87 mg glioblastoma cells, by American Type Culture Collection (1 mentions)
13 protocols using hek293t embryonic kidney cells
1
Stable Isotope-Labeled Nucleotide Synthesis
2
Characterization of Engineered Cell Lines
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HEK293T embryonic kidney cells (CRL‐3216), and 143B osteosarcoma cells (CRL‐8303) were obtained from ATCC. HEK293 Flp-In T-REx 293 cells were obtained from Invitrogen. The 143B.TK− rho0 derivative (143B206), obtained from Dr Carlos Moraes, was generated by Dr M.P. King (24 (link)). The details of the cell lines are listed in Supplementary Table S4 . HEK293T cells KO for MTG1, GTPBP10, or DDX28 were previously generated and reported by our group (4 ,9 (link),11 (link)). All cell lines were cultured at 37°C under 5% CO2 in high-glucose, sodium pyruvate, l -glutamine DMEM medium supplemented with 10% FBS and 100 μg/ml uridine (complete DMEM medium). Analysis of mycoplasma contamination was routinely performed. The chemicals and reagents used in this study are listed in the Supplementary Table S8 .
3
Culturing Human Cell Lines for Biomedical Research
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HEK293T embryonic kidney cells (CRL‐3216) and 143B osteosarcoma cells (CRL‐8303) were obtained from ATCC. The 143B.TK− rho0 derivative (143B206) was obtained from Dr M. King (31 (link)). The three cell lines were cultured at 37°C under 5% CO2 in high-glucose Dulbecco′s Modified Eagle′s Medium (DMEM) medium supplemented with 10% fetal bovine serum (FBS), 1 mM pyruvate and 50 μg/ml uridine (complete DMEM medium). Analysis for mycoplasma contamination was routinely performed.
iCell cardiomyocytes derived from induced pluripotent stem (iPS) cells were obtained from Cellular Dynamics (Madison, WI). These human heart cells consist of a 95% pure population of ventricular, atrial and nodal cells. They are known to recapitulate healthy human cardiac biology. Plating and maintenance media were obtained from Cellular Dynamics (https://cellulardynamics.com/products-services/icell-products/icell-cardiomyocytes/ ). Assays in cardiomyocytes are described in the Supplementary Data .
iCell cardiomyocytes derived from induced pluripotent stem (iPS) cells were obtained from Cellular Dynamics (Madison, WI). These human heart cells consist of a 95% pure population of ventricular, atrial and nodal cells. They are known to recapitulate healthy human cardiac biology. Plating and maintenance media were obtained from Cellular Dynamics (
4
Culturing HEK293T Cells
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HEK293T embryonic kidney cells (ATCC CRL-3216) were obtained from American Type Cell Culture Collection (ATCC) and maintained in Dulbecco’s modified Eagle’s medium (VWR, Cat# 45000-304). Culture media was supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin solution (CellGro, Cat# 30-002-CI). Cells were cultured at 37°C with 5% CO2 in a humidified incubator. Between passages, cells were detached with 0.25% trypsin with EDTA (VWR, Cat# 45000-664).
5
NSCLC and HEK293T Cell Lines
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The NSCLC cell lines A549, H460, H23, H1975, PC9, H1792, H1993, H1299, H2228, and HCC366, as well as HEK293T embryonic kidney cells, were purchased from ATCC (Manassas, VA, USA). HCC827 and HCC827/GR6 were kindly provided by Dr. Pasi A. Jänne (Dana-Farber Cancer Institute, Boston, MA, USA). All cells were maintained according to the providers’ instructions.
6
Culturing HEK293T and 143B Cell Lines
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HEK293T embryonic kidney cells (CRL-3216), and 143B osteosarcoma cells (CRL-8303) were obtained from ATCC. The 143B.TK− rho0 derivative (143B206) was obtained from Dr. M. King (35 (link)). The three cell lines were cultured at 37°C under 5% CO2 in high-glucose DMEM (Dulbecco's Modified Eagle Medium) supplemented with 10% FBS (fetal bovine serum), 1 mM pyruvate, and 50 μg/ml uridine (complete DMEM medium). Analysis for mycoplasma contamination was routinely performed.
7
Culturing MRC-5, HEK293T, and ARPE-19 Cells
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Human primary fetal lung fibroblasts (MRC-5, ATCC CCL-171), HEK293T embryonic kidney cells (ATCC CRL-3216) and ARPE-19 retinal pigment epithelial cells (ATCC CRL-2302) were purchased directly from ATCC, cultured in Thermo Fisher Gibco DMEM (MRC5, 293T) or DMEM/F12 (ARPE-19) full media supplemented with 10% (v/v) Fetal Bovine Serum (Cell Sera), 10 U/ml penicillin and 10 U/ml streptomycin (Thermo Fisher Gibco). Cells were maintained at 37°C and 5% CO2 in a humidified incubator and passaged 1:3 to 1:5 every third day for MRC5 and ARPE-19.
8
Cell Line Acquisition and Maintenance
9
Culturing Bladder Cancer and Kidney Cells
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J82 and T24 human bladder cancer cell lines and HEK 293T embryonic kidney cells were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA). The cells were cultured in RPMI-1640 medium and high-glucose Dulbecco’s modified Eagle medium (DMEM) supplemented with 10% fetal bovine serum (FBS) (Gibco™, Thermo Fisher Scientific, Waltham, MA, USA) and 1% penicillin-streptomycin (Gibco™, Thermo Fisher Scientific), respectively, in a humidified incubator with an atmosphere of 5% CO2 at 37 °C.
10
Glioma Patient Samples and Cell Lines
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Human surgical specimens were obtained from patients who underwent surgery at the Sun Yat-sen University Cancer Center (Guangzhou, China). This study was approved by the Institutional Ethical Review Board of Sun Yat-sen University Cancer Center (SL-B2023-039-01), and written informed consent was obtained from all patients. Final diagnoses of all the samples were made by our institutional neuropathologist (SR Liu) in accordance with the World Health Organization (WHO) criteria. The tumor tissues were snap frozen in liquid nitrogen and cryopreserved before use. A total of 50 glioma patient samples were included (Table 1 ).
Four human glioma cell lines (U87, U251, U138, and A172) were generously provided by Prof. Zhongping Chen (Sun Yat-sen University Cancer Center). They were cultured in DMEM (Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA). HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC) and cultured in DMEM supplemented with 10% FBS. All cells were incubated at 37 °C and 5% CO2 and tested to rule out mycoplasma contamination before use.
Four human glioma cell lines (U87, U251, U138, and A172) were generously provided by Prof. Zhongping Chen (Sun Yat-sen University Cancer Center). They were cultured in DMEM (Corning, NY, USA) supplemented with 10% fetal bovine serum (FBS, Gibco, Grand Island, NY, USA). HEK293T embryonic kidney cells were obtained from the American Type Culture Collection (ATCC) and cultured in DMEM supplemented with 10% FBS. All cells were incubated at 37 °C and 5% CO2 and tested to rule out mycoplasma contamination before use.
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