The plasmid expressing the surrogate reporter gene, pMRS-CMV (Toolgen) was transfected to HEK293T cells using Lipofectamine 3000 according to the manufacturer’s instructions. RFP-positive cells were sorted using Moflo Astrios EQ (Beckman Coulter), followed by selection and culture of cells with the highest RFP signals. CT26 murine colon cancer cells, 4T1 murine breast cancer cells, RENCA murine kidney cancer cells, Raw264.7 murine macrophage cells, and HEK293T human embryonic kidney cells were obtained from the American Type Culture Collection (ATCC). DCON murine gastric cancer cells were kindly provided by Dr. Sam S. Yoon (Memorial Sloan Kettering Cancer Center) and Dr. Sandra Ryeom (University of Pennsylvania) [42 (link)]. CT26, 4T1, and RENCA cells were cultured in RPMI-1640 (Gibco) supplemented with L-glutamine, 25 mM HEPES (Gibco), 10% fetal bovine serum (FBS, Hyclone) and 1% penicillin/streptomycin (P/S, Gibco). DCON, Raw264.7, HEK293T cells were cultured in DMEM (Gibco) containing the same supplements as RPMI 1640. Cells was cultured in a CO2 incubator (BB15, Thermo Fisher) at 37 ℃ at 5.0% CO2 condition.
Ct26 murine colon cancer cells
Manufactured by American Type Culture Collection
Sourced in United States
The CT26 murine colon cancer cells are a widely used cell line derived from a spontaneously generated murine colon adenocarcinoma. These cells are characterized by their ability to form tumors in immunodeficient mice.
Automatically generated - may contain errors
Lab products found in correlation
Raw 264.7 murine macrophage cells, by American Type Culture Collection (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
4t1 murine breast cancer cell, by American Type Culture Collection (1 mentions)
Moflo astrios eq, by Beckman Coulter (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Hek293t human embryonic kidney cells, by American Type Culture Collection (1 mentions)
Penicillin streptomycin p s, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Vero cell, by American Type Culture Collection (1 mentions)
293 cells, by American Type Culture Collection (1 mentions)
Penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Rpmi medium, by Thermo Fisher Scientific (1 mentions)
Balb c mice, by Janvier Labs (1 mentions)
4 protocols using ct26 murine colon cancer cells
1
Cell Culture and Transfection Protocol
2
Characterization of Cell Lines and Engineered Virus
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African green monkey kidney (Vero) cells, 293 cells and CT26 murine colon cancer cells were purchased from American Type Culture Collection (Manassas, VA, USA). 4T1 cells, a 6-thioguanine-resistant cell line derived from a BALB/c spontaneous mammary carcinoma [24 (link)], was kindly provided by Dr. Fred Miller (Michigan Cancer Foundation, Detroit, MI, USA). Cells were cultured with DMEM containing 10% fetal bovine serum (FBS). All media contained 100 U ml–1 penicillin and 100 mg ml–1 streptomycin (Invitrogen, Carlsbad, CA, USA). The details of the construction of the HSV-2-based FusOn-H2 have been reported [13 (link), 25 (link)]. FusOn-H3 was derived from FusOn-H2 by removing the GFP gene from the viral genome through homologous recombination. The virus was identified by picking a white plaque from the green background and GFP deletion was confirmed by DNA sequencing of the impacted region of the viral genome.
3
Culturing Pancreatic and Colon Cancer Cells
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BxPC-3 human pancreatic adenocarcinoma cells and CT26 murine colon cancer cells were purchased from the American Type Culture Collection (Manassas, VA), and RPMI 1640 medium containing 10% FBS and 1% penicillin-streptomycin was used as culture medium. Cells were cultured in a humidified atmosphere containing 5% CO2 at 37°C. Animals were obtained from Charles River Laboratories Japan Inc. (Yokohama, Japan). All the animal experiments were approved by the Animal Care and Use Committee of Tokyo Institute of Technology and the Animal Care and Use Committee of Kyoto University. The experiments were performed in accordance with the Guidelines for the Care and Use of Laboratory Animals as stated by Tokyo Institute of Technology and the Guidelines for the Care and Use of Laboratory Animals as stated by Kyoto University.
4
Subcutaneous CT26 Colon Cancer Model
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Tumors were established by subcutaneous injection of 3×105 CT26 murine colon cancer cells (CRL-2638, American Type Culture Collection, ATCC, VA, USA) suspended in 100 μL of culture medium over the thigh/flank of 7-week-old female BALB/c mice (Janvier Labs, France). Cells were grown in vitro in culture flasks using RPMI medium (Gibco, Thermo Fisher, Waltham, Massachusetts, USA) supplemented with bovine serum albumin (Gibco, Thermo Fisher, Waltham, Massachusetts, USA) and Penicillin Streptomycin (Pen Strep, (Gibco, Thermo Fisher, Waltham, Massachusetts, USA)) before inoculation in mice. Cells were tested for murine virus, human virus, mycoplasma, and lastly tested for cell line authentication to exclude cross-contamination during in vitro growth. Tumors were allowed to grow for 16 days before study start. The National Animal Experiments Inspectorate approved all study procedures (license no. 2016-15-0201-00920).
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