Hek293t

Manufactured by RIKEN BioResource Center

Sourced in Japan

HEK293T is a cell line commonly used in research laboratories. It is a variant of the HEK293 cell line, which was originally derived from human embryonic kidney cells. HEK293T cells are characterized by the expression of the SV40 large T antigen, which enables increased protein expression and higher transfection efficiency.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

HEK293T cells (23 citations) HEK293T cells (RBC2202) (2 citations)

20 protocols using hek293t

Genetic Manipulation of Cell Lines

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

S. albulus NBRC14147 was used to produce ε-PαL and its ester derivatives. E. coli BL21(DE3) was used to overexpress mAG, mKO, and Cre recombinase. Human cervical cancer cells (HeLa), human embryonic kidney cells (HEK293T), human hepatocellular carcinoma cells (HepG2), and human acute myelocytic leukemia cells (K562) were purchased from RIKEN BioResource Research Center (Japan). Human neuroblastoma cells (SH-SY5Y) and human dermal fibroblast cells (HDF) were purchased from the American Type Culture Collection (USA) and PromCell (Germany), respectively. The cell lines (HeLa, HEK293T, HepG2, K562) authenticated by RIKEN BioResource Research Center (Japan) were not further validated. The cell lines, SH-SY5Y and HDF, were not authenticated. We purchased the cell lines (HeLa, HEK293T, HepG2, K562) tested for mycoplasma contamination by RIKEN BioResource Research Center (Japan), but SH-SY5Y and HDF were not tested for mycoplasma contamination.
Two plasmids, pmAG1-S1 and pmKO1-MC1 (MBL, Japan), were used as the PCR templates to amplify the mAG and mKO genes, and the pmKO1-MC1 plasmid was also used to construct the Cre activity reporter plasmid. pKU470 (a gift from Dr. Haruo Ikeda)^{52 (link)} was used as a PCR template to amplify the Cre recombinase gene.

Takeuchi Y., Ushimaru K., Kaneda K., Maruyama C., Ito T., Yamanaka K., Ogasawara Y., Katano H., Kato Y., Dairi T, & Hamano Y. (2022). First direct evidence for direct cell-membrane penetrations of polycationic homopoly(amino acid)s produced by bacteria. Communications Biology, 5, 1132.

+ Open protocol

+ Expand

Cell Line Cultivation Protocols

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following cell lines were used in this study: HEK293T (a human embryonic kidney cell line stably expressing the SV40 large T antigen; RIKEN BioResource Center, Tsukuba, Japan), MCF-10A (a non-tumorigenic human breast epithelial cell line; ATCC, Rockville, MD), MCF-7 (a human breast cancer cell line; ATCC), SK-BR-3 (a human breast cancer cell line; ATCC), YMB-1-E (a human breast cancer cell line; JCRB Cell Bank, Tokyo), and MDA-MB-231 (a human breast cancer cell line; ATCC). MCF-10A cells were cultured in DMEM/Ham's F-12 (D/F) medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 100 ng/ml cholera toxin, 20 ng/ml epidermal growth factor (EGF), 0.01 mg/ml insulin, 500 ng/ml hydrocortisone and 5% FBS (Intergen, Purchase, NY). HEK293T, MCF-7, SK-BR-3, YMB-1-E and MDA-MB-231 cells were all cultivated in D/F medium (Thermo Fisher Scientific) supplemented with 10% FBS (Intergen).

Chen Y., Sumardika I.W., Tomonobu N., Kinoshita R., Inoue Y., Iioka H., Mitsui Y., Saito K., Ruma I.M., Sato H., Yamauchi A., Murata H., Yamamoto K.I., Tomida S., Shien K., Yamamoto H., Soh J., Futami J., Kubo M., Putranto E.W., Murakami T., Liu M., Hibino T., Nishibori M., Kondo E., Toyooka S, & Sakaguchi M. (2019). Critical role of the MCAM-ETV4 axis triggered by extracellular S100A8/A9 in breast cancer aggressiveness. Neoplasia (New York, N.Y.), 21(7), 627-640.

+ Open protocol

+ Expand

Culture and Maintenance of HT1080 and HEK293T Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human fibrosarcoma cell line HT1080 was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) and the human embryonic kidney (HEK) cell line HEK293T was obtained from RIKEN BioResource Center (Tsukuba, Japan). HT1080 and HEK293T cells were cultured in Dulbecco's modified Eagle's medium (DMEM; catalogue no. 05919; Nissui Pharmaceutical Co., Ltd., Tokyo, Japan) supplemented with 5% (v/v) fetal bovine serum (Bovogen Biologicals Pty Ltd., Melbourne, Australia), 100 mg/l kanamycin (Sigma-Aldrich, St. Louis, MO, USA), 100 U/ml penicillin G (Sigma-Aldrich), 600 mg/l L-glutamine (Sigma-Aldrich) and 2.25 g/l NaHCO₃ (Wako Pure Chemical Industries, Ltd., Osaka, Japan). Cells were incubated at 37°C in a humidified incubator with 5% CO2.

TSUCHIYA M., NIWA Y, & SIMIZU S. (2016). N-glycosylation of R-spondin1 at Asn137 negatively regulates its secretion and Wnt/β-catenin signaling-enhancing activity. Oncology Letters, 11(5), 3279-3286.

+ Open protocol

+ Expand

Cell Viability Assay on Cancer and Non-Cancer Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human cancer
cell lines, lung adenocarcinoma A549, multiple myeloma RPMI8226, and
glioma U87 were obtained from the American Tissue Culture Collection
(ATCC, Manassas, VA, USA). Human breast adenocarcinoma MDA-MB-231,
human embryonic kidney cells HEK-293T, and human vascular endothelial
EA.hy926 cell lines were purchased from Bioresource Collection and
Research Center (BCRC, Hsinchu, Taiwan). Cells were maintained in
a humidified incubator at 37 °C in 5% CO₂, and cell
viability was determined with the resazurin assay (Cayman Chemical,
USA) after the incubation with compounds for indicated concentrations
and times or DMSO as control.^{72 (link)}

Juang Y.P., Tsai J.Y., Gu W.L., Hsu H.C., Lin C.L., Wu C.C, & Liang P.H. (2024). Discovery of 5-Hydroxy-1,4-naphthoquinone (Juglone) Derivatives as Dual Effective Agents Targeting Platelet-Cancer Interplay through Protein Disulfide Isomerase Inhibition. Journal of Medicinal Chemistry, 67(5), 3626-3642.

+ Open protocol

+ Expand

Culturing Human and Mouse Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human lung adenocarcinoma cell line A549, mouse axillary lymph node/vascular epithelial cell line SVEC4-10 and human embryonic kidney cell line (HEK) 293T were obtained from the Bioresource Collection and Research Center of Taiwan. A549, SVEC4-10 and HEK293T cells were maintained in Dulbecco's modified Eagle medium (DMEM, Sigma-Aldrich) with 10% FBS at 37°C in humidified incubator with 5% CO₂.

Gu S.Y., Ho C.C., Huang Y.K., Chen H.W., Wang Y.C., Kuo C.Y., Teng S.C., Fu W.M., Yang P.C., Wu C.W., Peng F.C, & Ling T.Y. (2016). Acquisition of tumorigenic potential and enhancement of angiogenesis in pulmonary stem/progenitor cells through Oct-4 hyperexpression. Oncotarget, 7(12), 13917-13931.

+ Open protocol

+ Expand

Prostate Cell Culture Protocols

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human prostate fibroblast cells and fibroblast medium were purchased from ScienCell (Cat # 4430 and SC-2301; Carlsbad, CA, USA). The human normal prostate smooth muscle cells and PriGrow X medium were purchased from abm (cat #T4079 and TM4079, Richmond, BC, Canada). HEK-293T, PZ-HPV-7, CA-HPV-10, LNCaP, PC-3, and DU145 cells were obtained from the Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan) and cultured, as described previously [52 (link)]. The human prostate stromal myofibroblast WPMY-1 cells were purchased from the American Type Culture Collection (ATCC; CRL-2854; Manassas, VA, USA). The cells were cultured as instructed by the manufacturers. Fetal bovine serum (FBS) was purchased from HyClone Laboratories, Inc. (Logan, UT, USA); RPMI 1640 medium came from Life Technologies (Rockville, MD, USA) and Matrigel came from BD Biosciences (Bedford, MA, USA). TNFα and TGFβ were purchased from PeproTech (Cranbury, NJ, USA).

Chang K.S., Chen S.T., Sung H.C., Hsu S.Y., Lin W.Y., Hou C.P., Lin Y.H., Feng T.H., Tsui K.H, & Juang H.H. (2022). WNT1 Inducible Signaling Pathway Protein 1 Is a Stroma-Specific Secreting Protein Inducing a Fibroblast Contraction and Carcinoma Cell Growth in the Human Prostate. International Journal of Molecular Sciences, 23(19), 11437.

+ Open protocol

+ Expand

Transient Transfection of HEK293T Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The adherent human embryonic kidney (HEK) cell line HEK293T was obtained from Bioresource Collection and Research Center (BCRC, Taiwan) and cultured in α-MEM containing 100 U/mL penicillin/streptomycin and 10% fetal bovine serum at 37 °C, 5% CO₂ and 95% humidity. The HEK293T cells were transfected using Lipofectamine 2000 (Invitrogen) as described previously^{18 (link)}. Briefly, the ALPK1 full-length fragment was inserted into a plasmid vector (pHH-GM1, pEGFP-C2, pFN21K-Halo or pcDNA3.1), and Lipofectamine 2000 (Invitrogen) was used for transient transfection. The ratio of DNA to Lipofectamine 2000 was optimized for cell viability and transfection efficiency. The following ratio was used: 2.5 μg of DNA was combined with 10 μl of diluted Lipofectamine 2000 per 10 cm² of 80–90% confluent HEK293T cells. The plasmid DNA and Lipofectamine 2000 were separately diluted in opti-MEM and incubated for 5 min. They were combined along with serum-free α-MEM and incubated with cells for 5 h and then replaced with complete α-MEM. The cells were cultured for 24 h prior to harvest.

Lee C.P., Ko A.M., Chiang S.L., Lu C.Y., Tsai E.M, & Ko Y.C. (2018). Regulatory elements in vectors containing the ctEF-1α first intron and double enhancers for an efficient recombinant protein expression system. Scientific Reports, 8, 15396.

+ Open protocol

+ Expand

Mouse Primary Hepatocyte Isolation

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse primary hepatocytes were obtained from male KO or wild-type mice by the two-step collagenase perfusion method.^{42 (link)} Collagenase type II (Worthington Biochemical, USA; # CLS2)-digested liver was passed through a 70 mm cell strainer to obtain hepatocyte suspension. Then, mature hepatocytes were collected by centrifugation of the suspension. After isolation, hepatocytes were resuspended in DMEM supplemented with 5% FBS and 1% penicillin/streptomycin, and seeded on dishes coated by collagen type I (Cellmatrix type I-A, Nitta Gelatin Japan; #637–00653) at a density of 800,000cells/35mm dish. After over-night incubation under 5% CO2 at 95% humidity and 37°C, cells were used for experiments. HepG2 (Japanese Collection of Research Bioresources Cell Bank, Japan; JCRB1054) and HEK293T (RIKEN Bio Resource Research Center, Japan: RCB2202) were cultured in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin under 5% CO2 at 95% humidity and 37°C.

Xiao L., Alderisio K., Limor J., Royer M, & Lal A.A. (2000). Identification of Species and Sources of Cryptosporidium Oocysts in Storm Waters with a Small-Subunit rRNA-Based Diagnostic and Genotyping Tool. Applied and Environmental Microbiology, 66(12), 5492-5498.

+ Open protocol

+ Expand

Cell Culture of Transformed Human Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cell lines used were as follows: HEK293T (a human embryonic kidney cell line stably expressing the SV40 large T antigen; RIKEN BioResource Center, Tsukuba, Japan) and A549 (a human lung adenocarcinoma cell line with KRAS-mutant [G12S]; ATCC, Rockville, MD). These cells were all cultivated in D/F medium (ThermoFisher Scientific, Waltham, MA) supplemented with 10% fetal bovine serum (FBS).

Bajkowska K., Sumardika I.W., Tomonobu N., Chen Y., Yamamoto K.I., Kinoshita R., Murata H., Gede Yoni Komalasari N.L., Jiang F., Yamauchi A., Winarsa Ruma I.M., Kasano-Camones C.I., Inoue Y, & Sakaguchi M. (2020). Neuroplastinβ-mediated upregulation of solute carrier family 22 member 18 antisense (SLC22A18AS) plays a crucial role in the epithelial-mesenchymal transition, leading to lung cancer cells' enhanced motility. Biochemistry and Biophysics Reports, 22, 100768.

+ Open protocol

+ Expand

Cultivation of Human Lung and Kidney Cell Lines

Check if the same lab product or an alternative is used in the 5 most similar protocols

Human lung adenocarcinoma epithelial cell lines H1299 and A549 cells were obtained from American Type Culture Collection (ATCC, USA). Human embryonic kidney cells (HEK) 293 T was purchased from Bioresource Collection and Research Center (BCRC, Hsinchu, Taiwan). All of the three cell lines were maintained in Dulbecco’s Modified Eagle’s Medium (DMEM)/F12 (Thermo Fisher Scientific) supplemented with 10% FBS and 1% penicillin/streptomycin, and rested in a humidified incubator at 37 °C with 5% CO₂.

Hsieh K.Y., Tsai J.Y., Lin Y.H., Chang F.R., Wang H.C, & Wu C.C. (2021). Golden berry 4β-hydroxywithanolide E prevents tumor necrosis factor α-induced procoagulant activity with enhanced cytotoxicity against human lung cancer cells. Scientific Reports, 11, 4610.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!