Cells were cultured at 37 °C and 5% CO2(g). New cell lines were derived from a parent HEK293A cell line (ATCC) and cultured in Dulbecco’s modified Eagle’s medium (DMEM; Cellgro) supplemented with 10% fetal bovine serum (FBS; Cellgro), 1% penicillin-streptomycin (Cellgro), and 1% L-glutamine (Cellgro). For assays involving the tetracycline (Tet)-dependent transcriptional activation system (directed evolution of dox insensitivity, promoter activity assays, and reverse genetics), Tet-approved FBS (Takara Bio) was used. The producer and mutator cell lines (Table S2 ) were cultured in 50 μg/mL hygromycin (Thermo Fisher) to stably maintain transgenes, while the selector and phenotyping cell lines (Table S2 ) were cultured in 1 μg/mL puromycin (Corning) for the same purpose.
Hek293a cell line
Manufactured by American Type Culture Collection
Sourced in United States
The HEK293A cell line is a commonly used human embryonic kidney cell line. It is a subclone of the original HEK293 cell line. The HEK293A cell line is a versatile tool for various cell biology and biomedical research applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Corning (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Dulbecco modified eagle medium (dmem), by Corning (1 mentions)
Hygromycin, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Corning (1 mentions)
Puromycin, by Corning (1 mentions)
Tet approved fbs, by Takara Bio (1 mentions)
Pcr8 gw topo, by Thermo Fisher Scientific (1 mentions)
Ta cloning, by Thermo Fisher Scientific (1 mentions)
Du145, by American Type Culture Collection (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Lncap, by American Type Culture Collection (1 mentions)
Tissue culture treated flasks, by BD (1 mentions)
Dulbecco s modified eagle s medium dmem, by Caisson (1 mentions)
5 protocols using hek293a cell line
1
Culturing HEK293A Cell Lines for Genetic Studies
2
Construction of Adenoviral Vectors Encoding VHL and GFP
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Murine VHL gene (Gen Bank NM_009507.3) was obtained by RT-PCR using 5′-CCAATAATGCCCCGGAAGG-3′ (sense) and 5′-TCAAGGCTCCTCTTCCAGGTG-3′ (antisense) primers. cDNA were obtained from cellular RNA extracts of whole skin of healthy mice and were inserted into a Kosak (G/A)NNATGG sequence following insertion in the cloning vector. The GFP gene was amplified by PCR from commercial pEGFP plasmid (Invitrogen, Waltham, MA, USA). Both amplified products were cloned into the entry vector pCR®8/GW/TOPO® using TA cloning® (Invitrogen, Waltham, MA, USA), and were subsequently subcloned via recombination into the plasmid pAd/CMV/V5-DEST gateway (Invitrogen, Waltham, MA, USA). The recombinants were digested with the Pacl restriction enzyme (ThermoFisher, Waltham, MA, USA) to perform the transfection of the HEK-293A cell line (ATCC, Manassas, VI, USA). After 72 h of culture, the AdVHL and AdGFP vectors were harvested from the supernatant of transfected cells and kept in refrigeration for later amplification and purification. The AdLacZ was a control adenovirus provided by the manufacturer, and was included in cloning kit (Invitrogen, Waltham, MA, USA).
3
Cultivation and Maintenance of HEK293A and Capan-1 Cell Lines
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The HEK293A cell line was purchased from the American Type Culture Collection (ATCC) and cultured in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal calf serum. Capan-1 cells were a gift from Dr. Katharina Schlacher at MD Anderson Cancer Center and were grown in DMEM with 10% fetal bovine serum.
4
Culturing Human Cancer and Normal Cell Lines
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Prostate cancer cell lines DU145 and LNCaP, breast cancer cell line MDA-MB-231, lung cancer cell line A549, cervical cancer cell line HeLa, epidermoid carcinorma cell line A431, and human embryonic kidney (HEK) 293A cell line were purchased from the American Type Culture Collection (ATCC). Human foreskin normal fibroblast line Hs27 was purchased from UCSF Cell Culture Core Facility. Benign prostatic hyperplasia (BPH-1) cells were originally obtained from Dr. Gerald Cunha's lab at UCSF (10 (link)) and maintained in the lab. All cells were grown in high-glucose, l -glutamine, and sodium pyruvate-supplemented complete Dulbecco's modified Eagle's medium (DMEM) (Caisson Labs, North Logan, UT) with the addition of 10% fetal bovine serum (Fisher Scientific) and penicillin-streptomycin solution (Axenia BioLogix, Dixon, CA). Cells were grown in 5% CO2 at 37 °C on tissue culture-treated flasks (BD Biosciences). Cells were passaged utilizing 0.25% trypsin-EDTA (Invitrogen, Carlsbad, CA).
5
Cell Culture Protocols for HEK293A and Bovine Preadipocytes
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HEK293A cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). The bovine primary preadipocytes were isolated from the Qinchuan cattle and maintained at NBCIC (Wang Y. et al., 2018 (link)). HEK293A cells and bovine preadipocytes were cultured in DMEM-high glucose and DMEM-F12 (HyClone, Logan, UT, USA) medium, respectively, supplemented with 10% fetal bovine serum (FBS, Gibico, Carlsbad, CA, USA) and 1% penicillin-streptomycin (HyClone). The cells were incubated at 37°C under saturated humidity and 5% CO2.
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