Human BCa cell lines T24, RT‐112, and 253J were purchases from ATCC (Manassas, VA). T24 cells were cultured in McCoys 5A media with l ‐glutamine supplemented with 10% FBS and 1% penicillin/streptomycin sulfate and used within 6 months of purchase. RT‐112 and 253J cells were cultured in RPMI media with L‐glutamine supplemented with 10% FBS and 1% penicillin/streptomycin sulfate. All cells were tested for mycoplasma within the past 4 years (PCR Mycoplasma Detection Kit, MD Bioproducts). Cell lines were not authenticated after purchase.
Rt112
Manufactured by American Type Culture Collection
Sourced in United States
The RT112 is a laboratory equipment used for the cultivation and maintenance of cell cultures. It provides a controlled environment for the growth and propagation of various cell types. The RT112 unit maintains consistent temperature, humidity, and atmospheric conditions to support the optimal development of cultured cells.
Automatically generated - may contain errors
Lab products found in correlation
Umuc3, by American Type Culture Collection (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Sv huc 1, by American Type Culture Collection (4 mentions)
Biu 87, by American Type Culture Collection (4 mentions)
Tccsup, by American Type Culture Collection (3 mentions)
Rt112, by Thermo Fisher Scientific (3 mentions)
Um uc 3, by Thermo Fisher Scientific (3 mentions)
Bgj398, by Selleck Chemicals (3 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (3 mentions)
Ht1376, by American Type Culture Collection (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Hek293t, by American Type Culture Collection (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Ht1197, by American Type Culture Collection (2 mentions)
Ponatinib, by Selleck Chemicals (2 mentions)
Lapatinib, by Selleck Chemicals (2 mentions)
Acl 4 medium, by Thermo Fisher Scientific (2 mentions)
H1581, by Thermo Fisher Scientific (2 mentions)
26 protocols using rt112
1
Human BCa Cell Line Cultivation
2
Bladder and Colorectal Cancer Cell Lines
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Human bladder cancer cell lines HT1376, T24, UMUC3, RT112 and TCCSUP were originally from ATCC and were kindly provided by Drs. Yuesheng Zhang and Candace Johnson, Roswell Park Comprehensive Cancer Center. Dr. Johnson’s lab also provided human bladder cancer cell line 253 J B-V for this study. Human colorectal cancer (CRC) cell lines SNU-C2B and HCT117 were purchased from ATCC. Cells were grown under standard culture conditions at 37 °C in a humidified atmosphere containing 5% CO2. Cells were cultured in DMEM supplemented with 10% FBS, 100 unit/mL of penicillin and 100 μg/mL of streptomycin. Sub-confluent cells were treated with different concentrations of FL118 or with an equivalent amount of solvent DMSO according to the experiments.
3
Bladder Cancer Cell Line Protocol
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RT112, UMUC-3 (ATCC/LGC Promochem GmbH, Wesel, Germany) and TCCSUP (DSMZ, Braunschweig, Germany) bladder carcinoma cells were grown and subcultured in RPMI 1640, 10% fetal calf serum (FCS), 20 mM HEPES-buffer, 1% glutamax and 1% penicillin/streptomycin (all: Gibco/Invitrogen; Karlsruhe, Germany). RT112 is an invasive (pathological stage T2) moderately differentiated (grade 2/3) model of human bladder cancer, whereas TCCSUP is a transitional cell carcinoma, grade 4. UMUC-3 represents a high grade 3, invasive bladder cancer. Subcultures from passages 7–24 were used.
4
Bladder Cancer Cell Lines Cultivation
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The human urinary bladder transitional cell carcinoma cell lines T24 (RRID: CVCL_0554), UM-UC-3 (RRID: CVCL_1783) were purchased from ATCC, RT112 (RRID: CVCL_1670) was purchased from German Collection of Microorganisms and Cell Cultures GmbH (DSMZ) and UM-UC-1 (RRID: CVCL_2743) was purchased from European Collection of Authenticated Cell Cultures (ECACC). Human normal bladder epithelial cell line SV-HUC-1 (RRID: CVCL_3798) was purchased from ATCC. Murine transitional cell carcinoma cell line MB49 (RRID: CVCL_7076) was purchased from Millipore, and murine bladder epithelial cells (MBEC) were purchased from Procell (catalog no. CP-M058). Human lymphatic endothelial cells (HLEC) were obtained from ScienCell Research Laboratories. All cells were maintained in a humidified incubator with 5% CO2 at 37°C. The T24, RT112, and UM-UC-1 cells were cultured in RPMI 1640 medium (Gibco, catalog no. C11875500BT). The UM-UC-3 and MB49 cells were cultured in DMEM (Gibco, catalog no. C11995500BT). The SV-HUC-1 cells were cultured in Ham's F12K medium (Gibco, catalog no. 21127022). All medium was supplemented with 10% FBS (BI, catalog no. 04–001–1ACS). The HLECs were cultured in endothelial cell medium (ECM) with 5% FBS (ScienCell Research Laboratories, catalog no. 1001). The authentication and Mycoplasma testing of all cell lines were qualified.
5
Bladder Cancer Cell Line Culture
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Three epithelial bladder cancer cell lines, representing increasing malignancy states of progression, were used, RT112, T24 and J82 (ATCC, Rockville, MD). RT112 cells show an intermediate grade-2 and a stage Ta, i.e. these cells are moderately differentiated and have a small metastatic potential. T24 and J82 cells show a higher grade (grade-3) compared to RT112 cells and exhibit higher stages (T2–T3 and T3 respectively) i.e. they are poorly differentiated. J82 cells have the highest metastatic potential32 (link),52 (link).
Cancer cells were cultured in RPMI 1640 complete medium (Gibco, Saint Aubin, France) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The cell lines were stably transfected with the LifeAct GFP plasmid to stain F-actin53 (link). Cultures were grown at 37 °C in 5% CO2 atmosphere. Cells were let to grow until 50% confluency was reached. Then they were detached from culture dishes using trypsin-EDTA (0.25%), were re-suspended at cells/mL concentration in culture medium, and finally seeded in collagen gels.
Cancer cells were cultured in RPMI 1640 complete medium (Gibco, Saint Aubin, France) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin-streptomycin. The cell lines were stably transfected with the LifeAct GFP plasmid to stain F-actin53 (link). Cultures were grown at 37 °C in 5% CO2 atmosphere. Cells were let to grow until 50% confluency was reached. Then they were detached from culture dishes using trypsin-EDTA (0.25%), were re-suspended at cells/mL concentration in culture medium, and finally seeded in collagen gels.
6
Evaluating mRNA Targets in Urothelial Cancer
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To evaluate potential mRNA targets we studied 292 freshly frozen urothelial samples (Table 1 ) from two cohorts. The first was used to compare the expression of miRs-99a/100 and putative mRNA targets. The second examined these putative targets in a larger unrelated population. UCC were classified using the 2004 WHO/ISUP criteria and treated according to standard care [5 (link), 42 ]. Histologically normal urothelial samples were obtained from patients with UCC (distant to any tumor) and disease-free controls (at prostatectomy). We analyzed UCC cell lines representing the disease spectrum (RT4, RT112 and EJ/T24, respectively, purchased from ATCC) and normal non-immortalized human urothelial (NHU) cells [43 (link)].
7
Characterization of Human Cancer Cell Lines
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The human prostate cancer cell lines PC3 (docetaxel resistant, androgen-independent), DU145 (docetaxel sensitive, androgen-independent), LNCaP (docetaxel sensitive, androgen-dependent) [41 (link), 42 (link)], human bladder cancer cell lines RT112, RT4, 486p, T24, human embryonic kidney cell line HEK 293T, human embilical vascular endothelium cell line HUVEC, as well as human fibroblast cell lines MRC-5 and MRC-9 were obtained from ATCC (Manassas, VA, USA). The human germ cell tumor cancer cell line NCCIT was obtained from DSMZ (Braunschweig, Germany). TCam-2 and 2102EP cells were kindly provided by Prof. L. Looijenga (Rotterdam, The Netherlands). The cisplatin-resistant sublines NCCIT-R and 2102EP-R have been generated as reported before [16 (link), 17 (link)]. Cells were cultured according to the manufacturers instructions (culture conditions are described in the supplementary). Cells were continuously kept in culture for a maximum of 3 months, and were routinely inspected microscopically for stable phenotype and regularly checked for contamination with mycoplasma. Cell line authentication was performed by DSMZ (Braunschweig, Germany) using highly polymorphic short tandem repeat loci.
8
FGFR-Targeted Therapy in Cancer Cell Lines
9
Culturing Bladder Cancer Cell Lines
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T24 and RT-112, two bladder cancer cell lines, were purchased from ATCC and cultured in RPMI-1640 medium supplemented with 10% FBS.
10
Cell Culture Conditions for Cancer Research
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HCT116 (ATCC) cells were cultured in McCoy's 5A, RT112 and COLO205 (ATCC) cells were cultured in RPMI1640, and HEK 293 (ATCC) were cultured in DMEM media. Media were supplemented with 10% FBS (Sigma), 1% Pencillin/Streptomycin (Sigma) and 1% L-glutamine (Sigma). Cells were cultured at 37ºC and 5% CO 2 in a humidified incubator. Cells were STR authenticated, used between 3 and 15 passages, and tested for mycoplasma regularly (detection kit from Invivogen). HCT116-Cas9 cell line was generated by lentiviral transduction with lentiCas9-Blast (Addgene # 52962) and selection with 4 μg/mL of blasticidin (Sigma) for 3 days. KPC (Pdx1-cre;Kras LSL.G12D/+ ;Tp53 LSL.R172H/+ ) and KPCZ (Pdx1-cre;Kras ) cells are mouse pancreatic cancer cells without and with ZEB1 knockout, as previously described (28) , and were cultured in DMEM. TNFalpha was purchased from Gibco (Life technologies). Cdk inhibitor (CGP 60474) was purchased from Tocris (Biotechne).
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