Du145

Manufactured by American Type Culture Collection

Sourced in United States, United Kingdom, China, France, Germany, Sweden, Canada, Australia, Japan

The DU145 is a laboratory cell line derived from a human prostate carcinoma. It is widely used in cancer research for the study of cell biology and the development of potential therapies.

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1 453 protocols using du145

Cell Culture Protocol for Cancer Research

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The cell lines DU145 (HTB-81, human prostate carcinoma),
PC3 (CRL-1435,
human prostate adenocarcinoma), MCF7 (HTB-22, human breast adenocarcinoma),
MDA-MB231 (HTB-26, human breast adenocarcinoma), and HCT116 (CCL-247,
human colorectal carcinoma) were obtained from ATCC (American Type
Culture Collection, Manassas, VA, USA); WH1 human fibroblasts were
kindly provided by Dr. Guven.^{51 (link)} All the
cells were maintained under standard culture conditions (37 °C;
5% CO₂) in the RMPI1640 medium (PC3, MCF7, and MDA-MB231
cells), while DU145 and HCT116 cells were in the DMEM and Iscove’s
medium (WH1 cells) supplemented with 10% fetal calf serum, 1% glutamine,
and 1% antibiotics mixture; for HCT116 and DU145 cells, 1% sodium
pyruvate and 1% non-essential amino acids were also added to the culture
medium.

Fallica A.N., Barbaraci C., Amata E., Pasquinucci L., Turnaturi R., Dichiara M., Intagliata S., Gariboldi M.B., Marras E., Orlandi V.T., Ferroni C., Martini C., Rescifina A., Gentile D., Varchi G, & Marrazzo A. (2021). Nitric Oxide Photo-Donor Hybrids of Ciprofloxacin and Norfloxacin: A Shift in Activity from Antimicrobial to Anticancer Agents. Journal of Medicinal Chemistry, 64(15), 11597-11613.

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Cultivation of Prostate Cancer Cell Lines

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PC-3 and DU-145 human prostate cancer cells were acquired from ATCC (Rockville, MD, USA) and cultured in F12-K media (PC-3) purchased from Corning Incorporated (Oneonta, NY, USA) or Eagle’s Minimum Essential Medium (DU-145) purchased from ATCC. Media was supplemented with 10% fetal bovine serum and 1% antibiotic (streptomycin/penicillin). Cells were grown in incubator under standard conditions of 37 °C and 5% CO₂ through 10–25 passages.

Asay S., Graham A., Hollingsworth S., Barnes B., Oblad R.V., Michaelis D.J, & Kenealey J.D. (2020). γ-Tocotrienol and α-Tocopherol Ether Acetate Enhance Docetaxel Activity in Drug-Resistant Prostate Cancer Cells. Molecules, 25(2), 398.

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Cultivation of Prostate Cancer Cell Lines

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We obtained human prostate cancer PC3 (CRL‐1435), DU‐145 (HTB‐81), and LNCaP (CRL‐1740) cells from ATCC (Rockville, MD, USA). The PC3 cells were incubated in F‐12 K, 1X (Ham's F‐12 K Nutrient Mixture, Kaighn's Mod.) with L‐glutamine (10‐025‐CV) purchased from Corning Incorporated (Oneonta, NY, USA). The DU‐145 cells were incubated in Eagle's Minimum Essential Medium (30–2003) purchased from ATCC. The LNCaP cells were grown in RPMI‐1640 (30–2001) media purchased from ATCC. For each cell line, we added 10% fetal bovine serum (Corning, 35‐010‐CV) and 1% antibiotic (Hyclone, SV30010) and kept them in a 37°C humidified 5% CO2 incubator from passages 4–30. Docetaxel‐Resistant Clones were previously prepared in the lab as reported by Asay 2020.⁶

Berlin I.G., Jennings C.C., Shin S, & Kenealey J. (2023). Utilizing mixture design response surface methodology to determine effective combinations of plant derived compounds as prostate cancer treatments. Cancer Reports, 6(4), e1790.

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Prostate Cancer Cell Line Authentication

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The LNCaP, PC-3, 22Rv1, VCaP, and DU-145 human prostate cancer cell lines were purchased from American Type Culture Collection (ATCC). The identity of these cell lines has been authenticated by STR profiling, and all these cell lines tested negative for mycoplasma contamination. LNCaP (ATCC, CRL-1740) and 22Rv1 (ATCC, CRL-2505) cell lines were cultured with RPMI-1640 supplemented with 10% fetal bovine serum (FBS; PAN-Biotech, Bayern, Germany). PC-3 (ATCC, CRL-1435) was cultured with F-12K supplemented with 10% FBS (PAN-Biotech), VCaP (ATCC, CRL-2876), and DU145 (ATCC, HTB-81) cell lines were cultured with DMEM supplemented with 10% FBS (PAN-Biotech).

Wu M., Zhang R., Zhang Z., Zhang N., Li C., Xie Y., Xia H., Huang F., Zhang R., Liu M., Li X., Cen S, & Zhou J. (2023). Selective androgen receptor degrader (SARD) to overcome antiandrogen resistance in castration-resistant prostate cancer. eLife, 12, e70700.

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Cultivation of Prostate Cancer Cell Lines

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DU 145, LNCaP and PC-3 were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and conserved in liquid nitrogen at the Biological Resource, Jean Perrin Centre. Cells were cultivated in Eagle's minimum essential medium (EMEM) for DU 145 (ATCC), in RPMI 1640 medium for LNCaP (Gibco, Grand Island, NY, USA) and F-12K medium for PC-3 (ATCC). Cultures were supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Life Technologies, Carlsbad, CA, USA), 1% glutamine and 0.1% gentamicin (Panpharma, Luitré, France). Cells were maintained in a monolayer culture at 37° C in a humidified atmosphere of 95% air and 5% CO₂.

Daures M., Idrissou M., Judes G., Rifaï K., Penault-Llorca F., Bignon Y.J., Guy L, & Bernard-Gallon D. (2018). A new metabolic gene signature in prostate cancer regulated by JMJD3 and EZH2. Oncotarget, 9(34), 23413-23425.

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Cytotoxic Activity Evaluation of Cancer Cell Lines

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Human
cancer cell lines
A549 (lung adenocarcinoma, ATCC CCL-185), DU145 (androgen-human cancer
cell lines A549 (lung adenocarcinoma, ATCC CCL-185), MCF7 (breast
adenocarcinoma, ATCC HTB-22), DU145 (androgen-independent prostate
cancer, ATCC HTB-81), and HeLa (cervical carcinoma, ATCC CCL-2) were
obtained from ATCC. Human melanoma cell lines A375 (88113005) and
G-361 (88030401) were obtained from European Collection of Authenticated
Cell Cultures (ECACC). Cells were routinely grown in minimal essential
medium containing Eagle’s salts and l-glutamine (A549,
DU-145, MCF7, and HeLa) or Dulbecco’s Modified Minimal Essential
Medium (A375 and G-361) supplemented with 10% heat-inactivated fetal
bovine serum (FBS) in a humidified atmosphere of 5% CO₂ at 37 °C. To assess the antiproliferative activity of the compounds,
cells were seeded at a density of 2500 cells/well (HeLa), 5000 cells/well
(A549, DU145, and A375) or 10000 cells/well (MCF7 and G-361) in 96-well
plates, and cell viability was measured using the MTT assay. Values
are reported as the mean ± SD of two independent experiments.

Ortega J.A., Arencibia J.M., Minniti E., Byl J.A., Franco-Ulloa S., Borgogno M., Genna V., Summa M., Bertozzi S.M., Bertorelli R., Armirotti A., Minarini A., Sissi C., Osheroff N, & De Vivo M. (2020). Novel, Potent, and Druglike Tetrahydroquinazoline Inhibitor That Is Highly Selective for Human Topoisomerase II α over β. Journal of Medicinal Chemistry, 63(21), 12873-12886.

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Prostate Cell Line Culture Protocol

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Benign RWPE‐1 prostate epithelial cells (ATCC CRL‐11609) were cultured in in K‐SFM + Human recombinant Epidermal Growth Factor (rEGF) and Bovine Pituitary Extract (BPE) supplemented with 1% penicillin and streptomycin (all from Thermo Fisher Scientific). PCa cell line PC‐3 (CRl‐1435), VCap (CRL‐2876), DuCap (CVCL_2025), 22Rv1 (CRL‐2505), LNCaP (CRL‐1740) and DU145 (HTB‐81) were purchased from ATCC. VCaP and 293T‐D10 (a kind gift from Dr. Peppi Karppinen, University of Oulu) were grown in Dulbecco's Modified Eagle's Medium (DMEM) (11 965 092, Thermo Fisher Invitrogen), for DU145 Eagle's Minimum Essential Medium (EMEM) (30‐2003, ATCC) was used. LNCaP, VCaP and 22Rv1 were grown in Roswell Park Memorial Institute Medium (RPMI 1640) (R8758, Sigma). PC‐3 were grown in F12‐K (30‐2004, Invitrogen). DMEM, EMEM and RPMI were supplied with a final concentration of 10% fetal bovine serum (16 000 044, Thermo Fisher) and 1% of penicillin and streptomycin (15 140 122, Thermo Fisher). All cell lines were cultured in humidified incubators at 37 °C and 5% CO₂.

Cruz S.P., Zhang Q., Devarajan R., Paia C., Luo B., Zhang K., Koivusalo S., Qin L., Xia J., Ahtikoski A., Vaarala M., Wenta T., Wei G, & Manninen A. (2024). Dampened Regulatory Circuitry of TEAD1/ITGA1/ITGA2 Promotes TGFβ1 Signaling to Orchestrate Prostate Cancer Progression. Advanced Science, 11(11), 2305547.

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Characterization of Cell Lines in Cancer Research

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PC3, LNCaP, DU145, 22Rv1, VCaP, and SH-SY5Y cells lines were purchased from ATCC. KELLY cell line was purchased from Sigma-Aldrich. PC3-MYCN, LNCaP-ALK WT, LNCaP-YFP (Ctrl), LNCaP-ALK WT-dCas9, LNCaP-ALK WT-sgFAM150B, LNCaP-ALK WT-sgPTN, and LNCaP-ALK WT-sgNC cells were generated in this study. All cells were used within 10 passages of thawing and verified as mycoplasma free. PC3, LNCaP, DU145, 22Rv1, VCaP, and SH-SY5Y cell lines were genetically authenticated by ATCC. KELLY cell line was genetically authenticated by Public Health England.
For details, see Supplementary information.

Unno K., Chalmers Z.R., Pamarthy S., Vatapalli R., Rodriguez Y., Lysy B., Mok H., Sagar V., Han H., Yoo Y.A., Ku S.Y., Beltran H., Zhao Y, & Abdulkadir S.A. (2021). Activated ALK Cooperates with N-Myc via Wnt/β-catenin Signaling to Induce Neuroendocrine Prostate Cancer. Cancer research, 81(8), 2157-2170.

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Culturing Prostate Cancer Cell Lines

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LNCaP, DU145, PC3, and 22Rv1 PCa cell lines were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). Cells were cultured in a humidified incubator in 5% CO2 at 37 °C. DU145 and PC3 cells were cultured in RPMI-1640 medium (#30-2001, ATCC) supplemented with 10% Fetal Bovine Serum (FBS), (certified, One Shot™ format, Thermo Fisher Scientific, Waltham, MA, USA); 22Rv1 cells were cultured in Dulbecco’s modified Eagle’s Medium (DMEM) with 10% FBS; LNCaP cells were cultured in RPMI 1640 with 10% FBS supplemented with 10 mM HEPES (#ECM0180D, Euroclone, Mlan, Italy) and 1 mM Sodium Pyruvate (#ECM0542D, Euroclone, Mlan, Italy). All media were supplemented with antibiotics (150 U/mL penicillin, 200 U/mL streptomycin) (#ECB3001D, Euroclone, Mlan, Italy) and 2 mM Glutamine (#ECB3000D, Euroclone, Mlan, Italy). All cell lines were routinely tested using a PCR Mycoplasma Detection Set (#6601, Takara). Cell line authentication (STR) was carried out.

Cornice J., Capece D., Di Vito Nolfi M., Di Padova M., Compagnoni C., Verzella D., Di Francesco B., Vecchiotti D., Flati I., Tessitore A., Alesse E., Barbato G, & Zazzeroni F. (2021). Ultrasound-Based Method for the Identification of Novel MicroRNA Biomarkers in Prostate Cancer. Genes, 12(11), 1726.

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Prostate Cancer Cell Line Transfection Protocol

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RWPE-2 (CRL-11610), DU145 (HTB-81), VCaP (CRL-2876), PC-3 (CRL-1435) and LNCaP (CRL-1740) cells were purchased from American Type Culture Collection (ATCC). DU145, PC-3 and LNCaP cells were cultured in RPMI-1640 medium (ATCC^®30-2001™). RWPE-2 cells were cultured in K-SFM (ATCC^®17005-042™) and VCaP cells were cultured in Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc.). All cells were cultured in medium with fetal bovine serum (FBS; ATCC^®30-2020™; 10%) and gentamicin (Gibco; Thermo Fisher Scientific, Inc.; 50 mg/ml) at a density of 2x10⁴/ml in an incubator in an atmosphere with 5% CO₂ at 37˚C. After reaching 70% confluence, the cells were transfected with 10 nM miRNA-1297 inhibitor (Shanghai GenePharma, Inc.; 5'-CACCTGAATTACTTGAA-3'), NC inhibitor (Shanghai GenePharma, Inc.; 5'-ATACTCAAGCTTCTGAC-3') or PTEN-overexpression plasmid (Shanghai GenePharma, Inc.) using Lipofectamine 2000 (Invitrogen; Thermo Fisher Scientific, Inc.), according to the manufacturer's protocols. In brief, the mixture of Lipofectamine 2000 and miRNA-1297 inhibitor or PTEN plasmid was incubated for 10 min at room temperature. Next, the mixture was added into medium and incubated at 37˚C with cells for 48 h. Subsequently, cells were used for the follow-up experiments.

Wang L., Gao J., Zhang Y, & Kang S. (2021). Silencing miRNA-1297 suppresses the invasion and migration of prostate cancer cells via targeting modulation of PTEN and blocking of the AKT/ERK pathway. Experimental and Therapeutic Medicine, 22(1), 768.

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