Three BCa cell lines were used in this study, including T24, UM-UC-3, and TCCSUP (ATCC, Manassas, USA). T24 cells were cultured in Roswell Park Memorial Institute (RPMI)-1640 (Gibco, Shanghai, China), whereas UM-UC-3 and TCCSUP cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Gibco, Shanghai, China). All media were supplemented with 10% fetal bovine serum (FBS; Biological Industries, 04–001-1ACS) and 1% penicillin/streptomycin (Gibco, USA). The WDR5 inhibitor (OICR-9429) was purchased from Selleck (Houston, TX, USA) and dissolved in DMSO. Cells were grown in a 37 °C, 5% CO2 cell incubator with humidified atmosphere. The cells used in this study were excluded from mycoplasma contamination. Short tandem repeat (STR) authentication of the bladder cancer cell lines T24, UM-UC-3, and TCCSUP was conducted to prove that there was no misidentification or contamination with other cells. STR authentications were conducted by IGE Biotechnology LTD, Guangzhou, China.
Tccsup
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
The TCCSUP is a laboratory instrument designed for performing temperature control and cell culture applications. It serves as a regulated environment for maintaining optimal temperature conditions for cell growth and experimentation.
Automatically generated - may contain errors
Lab products found in correlation
Um uc 3, by Thermo Fisher Scientific (14 mentions)
Tccsup, by American Type Culture Collection (12 mentions)
Umuc3, by American Type Culture Collection (10 mentions)
Sv huc 1, by Thermo Fisher Scientific (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (9 mentions)
Sv huc 1, by American Type Culture Collection (9 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (7 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Mccoy s 5a medium, by Thermo Fisher Scientific (2 mentions)
Ham s f 12k kaighn s medium, by Thermo Fisher Scientific (2 mentions)
Sw780, by American Type Culture Collection (2 mentions)
Ht 1376, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Minimum essential medium (mem), by Thermo Fisher Scientific (2 mentions)
Biu 87, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Oicr 9429, by Selleck Chemicals (1 mentions)
F 12k media, by Thermo Fisher Scientific (1 mentions)
18 protocols using tccsup
1
Bladder Cancer Cell Line Cultivation
2
Culturing Poorly Differentiated Bladder Cancer Cell Lines
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Two poorly differentiated bladder cancer cell lines T24 and TCCSUP were purchased from ATCC. These cell lines were grown in cell culture media (T24 in McCoy's 5a medium modified, Gibco: 16600082; TCCSUP in Minimum essential medium Gibco: 11095080 and 1 mM sodium pyruvate) added with 10% fetal bovine serum, 1% penicillin/streptomycin.
3
Bladder Cancer Cell Lines and Tissue Samples
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The normal human bladder cell line SV-HUC-1 and human BC-derived cell lines (5637, T24, TCCSUP, SW780, J82, and UM-UC-3) were obtained from the American Type Culture Collection (Manassas, VA, United States). The SV-HUC-1 cell line was cultured in Ham’s F-12K (Kaighn’s) medium (Gibco). The 5637 cell line was cultured in RPMI-1640 medium (Gibco). The T24 cell line was cultured in McCoy’s 5A medium (Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco). The TCCSUP, J82 and UM-UC-3 cell lines were cultured in minimum essential medium (Gibco). All media mentioned above were supplemented with 10% FBS (Gibco), 100 U/ml penicillin and 100 U/ml streptomycin. Cells were incubated in a humidified atmosphere containing 5% CO2 in air at 37°C. All tissue specimens including 30 BC tissues and 30 adjacent normal tissues for RT-qPCR were obtained from BC patients without undergoing radiotherapy and chemotherapy diagnosed at the Department of Urology, Peking University Shenzhen Hospital, China. All experiments followed the “Helsinki Declaration” and were approved by the Ethics Committee of Peking University Shenzhen Hospital. All patients were informed of their specimens content, potential risks, purpose and signed written informed consent.
4
Cell Lines and Actinomycin D Assay
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Human BC cell lines T24, UM-UC-3, RT4, J82, 5637, HT-1376, TCCSUP and the immortalized normal uroepithelium cell line SV-HUC-1 were purchased from American Type Culture Collection (ATCC, USA). T24 and 5637 cells were cultured in RPMI-1640 (Gibco, Shanghai, China), RT4 cells were cultured in McCoy’s 5A(Gibco), UM-UC-3, J82, HT-1376, TCCSUP and HEK-293 T cells were cultured in DMEM (Gibco), whereas SV-HUC-1 cells were maintained in F-12 K media (Gibco), supplemented with 10% FBS (Gibco, USA) and 1% penicillin/streptomycin (Gibco). Cells were grown in a humidified atmosphere of 5% CO2 at 37 °C. For Actinomycin D assay, T24 and UM-UC-3 cells were exposed to 2 μg/mL Actinomycin D (Sigma, USA) to block transcription for 8, 16 and 24 h.
5
Culturing Bladder Cancer and HUVEC Cells
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SV-HUC-1, BUC cell lines (T24, UM-UC-3, 5637, J82 and TCC-SUP) and HUVEC were purchased from the American Type Culture Collection (Manassas, Virginia, USA). Cells were cultured in DMEM (SV-HUC-1, UM-UC-3, T24 and HUVEC), McCoy’s 5A (J82) and RPMI-1640 (5637 and TCC-SUP) basal medium (Gibco, Gaithersburg, MD, USA), which were supplemented with 10% fetal bovine serum (FBS, MilliporeSigma, Burlington, MA, USA), 100 U/ml penicillin and 0.1 mg/ml streptomycin (Beyotime, Beijing, China). Cells were incubated at 37 °C in 5% CO2 incubator. The medium was changed every 1–3 days.
6
Bladder Cancer Cell Line Culturing
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Human bladder cancer cell lines J82, TCCSUP, EJ, UMUC3, T24, 5637 and RT4, human immortalized uroepithelium cell line SV-HUC-1, were purchased from American Type Culture Collection (ATCC, USA). The human bladder cancer cell line T24T was provided by Dr. Dan Theodorescu (Departments of Urology, University of Virginia, Charlottesville, VA, USA) as described in our previous studies [24 (link)]. J82 and TCCSUP cells were cultured in MEM (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin/streptomycin (Gibco, USA). T24T and UMUC3 cells were cultured in DMEM (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin/streptomycin (Gibco, USA). 5637, EJ, T24 and RT4 cells were cultured in RPMI-1640 medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin/streptomycin (Gibco, USA). SV-HUC-1 were cultured in F-12K medium (Gibco, USA) supplemented with 10% FBS (Gibco, USA), 1% penicillin/streptomycin (Gibco, USA). Cells were cultured in an incubator at 37 °C with humidifified atmosphere of 5% CO2. All bladder cancer cell lines were confirmed within 6 months before use by using a short tandem repeat profiling and were confirmed negative for Mycoplasma contamination.
7
Bladder Cancer Cell Line Cultivation
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Bladder cancer cell lines (T24, 5637, BIU-87, UM-UC-3, RT4, EJ, TCCSUP) and uroepithelial cell SV-HUC-1 was purchased from the Cell Bank of the Chinese Academy of Sciences. Primary normal bladder urothelial cells (NBUCs) cultures and human uroepithelial cell SV-HUC-1 were established as described previously [14 (link)]. The cell lines EJ, 5637, BIU-87, UM-UC-3, T24, and TCCSUP were maintained in RPMI 1640 medium, while the RT4 cell line was cultured in McCoy's 5a medium supplemented with 10% fetal bovine serum (FBS) (Gibco). SV-HUC-1 cells was grown in F12K medium supplemented with 10% FBS.
8
Bladder Cancer Cell Lines Cultivation
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Human BLCA cell lines (BIU-87, 5637, EJ, T24, and TCCSUP) and normal urothelial cell line SV-HUC-1 were purchased from National Infrastructure of Cell Line Resource (Beijing, China). BIU-87, 5637, and EJ cell lines were maintained in Roswell Park Memorial Institute 1640 medium (Gibco, Carlsbad, California, USA) encompassing 10% fetal bovine serum (Gibco) and penicillin/streptomycin (Gibco).
T24 and TCCSUP cell lines were cultured in complete Dulbecco’s Modified Eagle Media (Gibco), and SV-HUC-1 cell line was incubated in complete Ham’s F12K (Kaighn’s) modified medium (Gibco). All cells were cultured in a humidified CO2 condition at 37 °C.
T24 and TCCSUP cell lines were cultured in complete Dulbecco’s Modified Eagle Media (Gibco), and SV-HUC-1 cell line was incubated in complete Ham’s F12K (Kaighn’s) modified medium (Gibco). All cells were cultured in a humidified CO2 condition at 37 °C.
9
Bladder Cancer Tissue and Cell Line Protocol
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Human BC samples (60 cases) with the matched normal tissues (10 cases) as controls were obtained from patients who underwent radical cystectomy at the Jiangxi Provincial People’s Hospital Affiliated to Nanchang University from January 2012 to December 2016. This study and the written informed consents obtained from all patients were approved by the Ethics Committee and the Institutional Review Board of the Jiangxi Provincial People’s Hospital and conducted in compliance with the Declaration of Helsinki. The histopathology of tissue samples was determined and confirmed by two pathologists according to the criteria of the WHO and the Nevin stage system. Human BC cell lines T24, HT1376, TCCSUP, and immortalized human bladder cell SV-HUC-1 were purchased from China Academia Sinica Cell Repository (Shanghai, China), T24 was cultured in RPMI 1640 supplemented with 10% FBS(GIBCO, USA),while HT1376 and TCCSUP were cultured in DMEM (GIBCO, Grand Island, NE, USA), and SV-HUC-1 was cultured in F12K (GIBCO, USA) and all were incubated in incubator at 37°C with a 5% CO2 atmosphere. T24 was treated with 10 μM SP600125 (Selleck Chemicals, Houston, TX, USA) for 1 hr to inhibit the JNK pathway.
10
Cell Culture Maintenance Protocol
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All cell lines were obtained from the American Type Culture Collection and maintained using standard media and conditions. Human BC cells (T24, TCCSUP, UM-UC-3, J82 and 5637), human normal bladder epithelial cells (SV-HUC-1) and 293T cells were maintained in Roswell Park Memorial Institute (RPMI)-1640 medium, Dulbecco's modified Eagle's medium (DMEM) or F-12K supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin (PS) (all from Gibco; Thermo Fisher Scientific Inc.). All cells were cultured at 37°C in a 5% CO2 incubator.
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