HeLa cells were purchased from the Human Science Research Resources Bank (Sennanshi, Japan). HEK-293T cells were obtained from Invitrogen as Lenti-X 293 cells (Carlsbad, CA). A549 cells were obtained from the American Tissue Culture Collection. HeLa, HEK-293T, and A549 cells were maintained in DMEM (Wako, Osaka, Japan) supplemented with 10% FBS. For imaging, these cells were plated on 35-mm glass base dishes (Asahi Techno Glass, Tokyo, Japan). One hour before observation, HeLa and A549 cells were maintained with phenol red-free Medium 199 (Invitrogen) containing 0.1% bovine serum albumin and 20 mM HEPES.
Hek293t
Manufactured by Fujifilm
Sourced in Japan
HEK293T is a cell line derived from human embryonic kidney cells. It is commonly used in laboratory research as a host cell for the production and study of various proteins, viruses, and other biological materials.
Automatically generated - may contain errors
Lab products found in correlation
Hek293t, by American Type Culture Collection (5 mentions)
Dulbecco modified eagle medium (dmem), by Fujifilm (4 mentions)
Penicillin streptomycin, by Fujifilm (2 mentions)
Rpmi 1640 medium, by Fujifilm (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Hek293t cells, by Thermo Fisher Scientific (1 mentions)
A549 cell, by American Type Culture Collection (1 mentions)
Nhdfs, by Takara Bio (1 mentions)
Capan 1, by Fujifilm (1 mentions)
Iscove modified dulbecco media (imdm), by Fujifilm (1 mentions)
Miapaca 2, by Fujifilm (1 mentions)
Fetal bovine serum (fbs), by Fujifilm (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Fujifilm (1 mentions)
Capan 1, by American Type Culture Collection (1 mentions)
Mia paca 2, by American Type Culture Collection (1 mentions)
Antibiotic antimycotic mixed solution, by Nacalai Tesque (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Renca, by American Type Culture Collection (1 mentions)
6 protocols using hek293t
1
Cell Culture and Imaging Protocol
2
PDAC and HEK293T Cell Culture Protocol
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The human PDAC cell lines Capan-1 and MIAPaCa-2 and the embryonic kidney cell line HEK293T were purchased from the American Type Culture Collection (ATCC, Rockville, MD, USA). NHDFs were purchased from TAKARA BIO (Shiga, Japan). The PCI43P5 cell line was previously established from surgically resected PDAC tissues at our institute [32 (link)]. All cell lines were cultured in the appropriate medium as directed by the manufacturers or according to published studies. Capan-1 was cultured in IMDM (WAKO, Tokyo, Japan) with 20% fetal bovine serum (Cell Culture Bioscience), and 1% penicillin/streptomycin (Life Technologies), and MIAPaCa-2, PCI43P5, NHDF, and HEK293T cells were cultured in RPMI 1640 (WAKO) with 10% fetal bovine serum, and 1% penicillin/streptomycin. All cells were incubated at 37°C in a mixture of 5% CO2 and 95% air.
3
Cell Culture of C2C12 and HEK293T
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Mouse C2C12 myoblast and human embryonic kidney 293T (HEK293T) cell lines were purchased from American Type Culture Collection (Manassas, VA, USA). C2C12 myoblasts and HEK293T cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM; FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) with 10% fetal bovine serum (FBS; Sigma-Aldrich, Tokyo, Japan) and an antibiotic/antimycotic mixed solution (100 U/mL penicillin G, 100 µg/mL streptomycin, and 0.25 µg/mL amphotericin B) (Nacalai Tesque). Cells were cultured in a humidified atmosphere at 5% CO2 and 37 °C; the medium was replaced every two days.
4
Cell Culture and Reagent Preparation
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The HEK293T (human embryonic kidney cell), CT26 (murine colon cancer), RENCA (murine renal cell carcinoma), and A20 (murine lymphoma) cell lines were purchased from ATCC (Manassas, VA). The HEK293T and CT26 cell lines were maintained in DMEM (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan) supplemented with 10% foetal calf serum (FCS; Cytiva, Tokyo, Japan). The RENCA and A20 cell lines were maintained in RPMI medium (FUJIFILM Wako Pure Chemical Corporation) supplemented with 10% FCS. All tumour cells were confirmed to be Mycoplasma (−) using a PCR Mycoplasma Detection Kit (TaKaRa, Shiga, Japan) according to the manufacturer’s instructions before use. Recombinant human Wnt3a protein was obtained from R&D (Minneapolis, MN). Recombinant murine Wnt3a protein was purchased from PeproTech (Cranbury, NJ). Rat anti-mouse PD-1 mAb (RMP1-14) and control rat IgG2a mAb (RTK2758) were obtained from BioLegend (San Diego, CA).
5
Diverse Cell Lines for Cancer Research
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B16 (murine melanoma; F1 (parental), F10 (metastatic)), HEK293T (human embryonic kidney), ES2 (human clear cell ovarian carcinoma), and MOLM14 (human acute myeloid leukemia) cell lines were obtained from the American Type Culture Collection (ATCC, Manassas, VA, USA). The MC38 (murine colon adenocarcinoma) cell line was procured from Kerafast. The cells were cultured in either RPMI-1640 medium (FujiFilm Wako Pure Chemical, Osaka, Japan) for HEK293T, MOLM14, and K562 cells or DMEM (FujiFilm Wako Pure Chemical) for B16F1, B16F10, MC38, and ES2 cells supplemented with 10% heat-inactivated fetal bovine serum (FBS, Biosera, Cholet, France) and antibiotics (100 U penicillin/mL and 100 μg streptomycin/mL, Thermo Fisher Scientific, Waltham, MA, USA) at 37°C in a humidified atmosphere containing 5% CO2.
6
Cell Culture and Transfection Protocol
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The human cell lines used in this study include HCT116, DLD-1, AsPC-1, PL5 and HEK293T (American Type Culture Collection, Manassas, VA, USA) and HuP-T3 (Sigma-Aldrich, St Louis, MO, USA). Cell culture was carried out in McCoy's 5A modified medium (Sigma-Aldrich; for HCT116), RPMI-1640 medium (Wako, Osaka, Japan; for DLD-1, AsPC-1 and PL5) and Dulbecco's modified Eagle's medium (Wako; for HuP-T3 and HEK293T). Media were supplemented with 5% fetal bovine serum (BioWest, Nuaillé, France) and 1% penicillin/streptomycin (Wako) prior to use for regular cell culture. The transfection of cells with the plasmids was performed using TransIT-LT1 transfection reagent (Mirus Bio, Madison, WI, USA) according to the manufacturer's instructions. Selection of cells with G418 (Life Technologies) was carried out at concentrations of 0.4 mg/ml (PL5 and HuP-T3), 0.8 mg/ml (HCT116 and AsPC-1) or 1 mg/mL (DLD-1) throughout the study.
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