The prostate cancer cell lines PC-3 (androgen-insensitive) and LNCaP (androgen-sensitive) and the histologically normal prostate epithelial RWPE-1 cell line (immortalized with papilloma virus 18) were purchased from Sigma-Aldrich (Barcelona, Spain). PC-3 cells were cultured in Ham’s F-12K (Kaighn’s) Medium (1:1 mixture) with L-glutamate (Invitrogen/Gibco, Fisher Scientific SL, Madrid, Spain). LNCaP cells were cultured in RPMI 1640 medium (Merck KGaA, Darmstadt, Germany) supplemented with 1-mM sodium pyruvate (Gibco). PC-3 and LNCaP cultures were supplemented with 10% fetal bovine serum. RWPE-1 cells were cultured in keratinocyte serum-free medium plus 5 μg/mL bovine pituitary extract. All the cells were supplemented with 1× antibiotic-antimycotic (Gibco) and 5 μg/mL PlasmocinTM (InvivoGen, San Diego, CA, USA). Where indicated, the cells were grown in serum-deprived medium overnight before stimulation for 24 or 48 h with 100 ng/mL human recombinant TWEAK (PeproTech, bioNova cientifica, Barcelona, Spain) and were cultured in a humidified 5% CO2 atmosphere at 37 °C.
Lncap
Manufactured by Merck Group
Sourced in United States, United Kingdom, Germany, Spain
LNCaP is a cell line derived from a human prostate cancer sample. It is commonly used in cancer research and drug development. The core function of LNCaP is to serve as an in vitro model for studying prostate cancer biology and evaluating potential therapeutic agents.
Automatically generated - may contain errors
Lab products found in correlation
Lncap, by American Type Culture Collection (45 mentions)
Du145, by American Type Culture Collection (22 mentions)
Rpmi 1640 medium, by Merck Group (20 mentions)
Du145, by Merck Group (19 mentions)
Rpmi 1640, by Merck Group (13 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (12 mentions)
Fetal bovine serum (fbs), by Merck Group (10 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (10 mentions)
Rwpe 1, by American Type Culture Collection (8 mentions)
Penicillin, by Merck Group (7 mentions)
Streptomycin, by Merck Group (7 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (7 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (6 mentions)
L glutamate, by Thermo Fisher Scientific (5 mentions)
Streptomycin, by Thermo Fisher Scientific (5 mentions)
Plasmocin, by InvivoGen (4 mentions)
Poly dl ornithine, by Merck Group (4 mentions)
Penicillin streptomycin, by Merck Group (4 mentions)
L glutamine, by Merck Group (4 mentions)
Lncap, by Thermo Fisher Scientific (4 mentions)
71 protocols using lncap
1
Prostate Cancer Cell Line Culture Protocols
2
Transwell Co-culture of PPAT Explants and PCa Cells
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We designed Transwell co-culture assays of PPAT explants (7 low-risk and 3 high-risk) with 2 PCa cell lines. The prostate cancer cell lines PC-3 (androgen-insensitive) and LNCaP (androgen-sensitive) were purchased from Sigma-Aldrich (Barcelona, Spain). PC-3 cells were cultured in Ham’s F-12 K (Kaighn’s) Medium (1:1 mixture) with L-glutamate (Invitrogen/Gibco, Fisher Scientific SL, Madrid, Spain). LNCaP cells were cultured in RPMI 1640 medium (Merck KGaA, Darmstadt, Germany) supplemented with 1 mM sodium pyruvate (Gibco). PC-3 and LNCaP cultures were supplemented with 10% fetal bovine serum, 1% penicillin/streptomycin, and 5 μg/mL Plasmocin® (Invivogen, IBIAN Technologies, Zaragoza, Spain). Cells were seeded into Transwell 0.4-μm pore size cell culture inserts (Fisher scientific, Barcelona, Spain) at 50,000 cells/0.9 cm2 in the same medium at 37 °C and 5% CO2 during 24 h. The next day, the medium was exchanged for serum-free medium, for 24 h. Subsequently, 50 mg of fresh PPAT was washed with PBS twice and added to the lower Transwell chamber in 1 mL of M199 medium with 10% FBS and 1% penicillin/streptomycin in 25 mM HEPES. Each sample was tested in duplicate. The co-culture was maintained at 37 °C and 5% CO2 for 48 h in the same medium. Subsequently, cells and tissue explants were removed, and RNA was extracted.
3
Profiling Prostate Cancer Cell Lines
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Following commercial prostate cells, representing different stages of prostate cancer, were used in the present study (Supplementary File 1); PNT2, P4E6 and LNCaP (Sigma Aldrich, St. Louis, MO, USA), DU145 and PC3 (ATCC, Manassas, VA, USA). PNT2 and LNCaP cells were cultured in RPMI-1640 supplemented with 10% FBS and 1% PEST (Sigma Aldrich), whereas P4E6 cells were maintained in Stemline Keratinocyte Medium II with Stemline Keratinocyte Growth Supplement, 2 mM Glutamine and 2% of FBS (Sigma Aldrich), DU145 cells were cultured in EMEM (Sigma Aldrich) supplemented with 10% FBS and 1% PEST and PC3 cells were cultured in DMEM (Sigma Aldrich) containing 10% FBS, 5% of pyruvate sodium (Sigma Aldrich) and 1% PEST.
4
Prostate Cancer Cell Line Characterization
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The human prostate carcinoma cell lines LNCaP (RRID: CVCL_1379), VCaP (RRID: CVCL_2235), PC3 (RRID: CVCL_0035), 22Rv1 (RRID: CVCL_1045) and C4-2B (RRID: CVCL_4784) were obtained from the ATCC. LNCaP-V16D, LNCaP-MR49F, and CWR-R1-D567 have been described previously (16, 17 ). Luciferase-labelled RM1, a murine syngeneic model of bone-metastatic prostate cancer, has been described previously (18 (link)). C4-2B, 22Rv1, LNCaP, and LNCaP-V16D cells were maintained in RPMI1640 (Sigma-Aldrich) containing 10% FBS and 2 mmol/L l -Glutamine. PC3 cells were cultured in RPMI1640 containing 5% FBS and 2 mmol/L l -Glutamine. LNCaP-MR42D and LNCaP-MR49F were maintained in RPMI1640 containing 10% FBS, 10 μmol/L enzalutamide, and 2 mmol/L l -Glutamine. CWR-R1-D567 cells were maintained in RPMI1640 containing 10% charcoal-stripped serum and 2 mmol/L l -Glutamine. VCaP cells were maintained in DMEM (high glucose) containing 10% FBS, 2 mmol/L l -Glutamine, 2 mmol/L sodium pyruvate, and 2 mmol/L of nonessential amino acids solution (Sigma-Aldrich). All cell lines were authenticated by short tandem repeat profiling by CellBank Australia in 2017–2020 and were regularly screened for potential Mycoplasma contamination.
5
Lentiviral Knockdown of Cx43 in Prostate Cancer
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Prostate cancer cell lines LNCaP, C4-2, DU145, PC-3 were maintained in RPMI medium supplemented with 10% fetal bovine serum (FBS), 50 U/ml of penicillin, 50 μg/ml of streptomycin and 2 mM L-glutamine (Life Technologies). LNCaP was purchased from ATCC, C4-2 was kindly provided by Dr. Alex Almasan (Cleveland Clinic), and DU145 and PC-3 were kindly provided by Dr. Nima Sharifi (Cleveland Clinic). LNCaP and PC-3 cells with down-regulated Cx43 were prepared by transduction with lentivirus expressing Cx43 specific shRNA (Sigma, TRCN0000059773 and TRCN0000059775). Lentivirus was produced in 293T cells using standardized protocol with packaging plasmids as described previously [33 (link)], and viral particles containing conditioned medium was filtered through 0.45 μM PVDF membrane and directly used to infect prostate cancer cell lines in the presence of 8 μg/ml polybrene (Sigma). Twenty four hours after infection, cells were selected with puromycin (Sigma) at a final concentration of 3 μg/mL for PC-3 cells and 1 μg/mL for LNCaP cells. Surviving cells were pooled together and maintained in medium containing 1 μg/mL puromycin for both cell lines. Control cells were obtained in a similar manner with lentivirus expressing non-targeting shRNA (Sigma, SHC002).
6
Cell Line Sourcing and Maintenance
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Tumor cell lines OVCAR-3, SKOV-3, HT-29, and HeLa were obtained from ATCC. LNCaP and MCF-7 were obtained from Millipore Sigma. IGROV-1 was obtained from NCI. Choroid plexus, retinal pigment, pulmonary alveolar, bronchial, and renal cortical epithelial cells were obtained from ScienCell Research Laboratories. All cell lines were maintained in a 5–8% CO2 buffered incubator at 37°C and maintained as per manufacturer’s recommendations.
7
Developing Novel Proteasome Inhibitors for Cancer Treatment
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Example 8
New proteasome inhibitors may be developed not only for treating conditions mediated by senescent cells, but also conditions mediated by cancer cells.
The ability of compounds to specifically kill cancer cells can be tested in assays using other established cell lines. These include HeLa cells, OVCAR-3, LNCaP, and any of the Authenticated Cancer Cell Lines available from Millipore Sigma, Burlington Mass., U.S.A. Compounds specifically kill cancer cells if they are lethal to the cells at a concentration that is at least 5-fold lower, and preferably 25- or 100-fold lower than a non-cancerous cell of the same tissue type. The control cell has morphologic features and cell surface markers similar to the cancer cell line being tested, but without signs of cancer.
In vivo, compounds are evaluated in flank xenograft models established from sensitive SCLC (H889) and hematologic (RS4; 11) cell lines, or using other tumor-forming cancer cell lines, according to what type of cancer is of particular interest to the user. When dosed orally or intravenously, compounds induce rapid and complete tumor responses (CR) that are durable for several weeks after the end of treatment in all animals bearing H889 (SCLC) or RS4; 11 (ALL) tumors. Similar treatment of mice bearing H146 SCLC tumors can induce rapid regressions in the animals.
8
Evaluating Novel Proteasome Inhibitors for Cancer Treatment
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Example 8
New proteasome inhibitors may be developed not only for treating conditions mediated by senescent cells, but also conditions mediated by cancer cells.
The ability of compounds to specifically kill cancer cells can be tested in assays using other established cell lines. These include HeLa cells, OVCAR-3, LNCaP, and any of the Authenticated Cancer Cell Lines available from Millipore Sigma, Burlington Mass., U.S.A. Compounds specifically kill cancer cells if they are lethal to the cells at a concentration that is at least 5-fold lower, and preferably 25- or 100-fold lower than a non-cancerous cell of the same tissue type. The control cell has morphologic features and cell surface markers similar to the cancer cell line being tested, but without signs of cancer.
In vivo, compounds are evaluated in flank xenograft models established from sensitive SCLC (H889) and hematologic (RS4;11) cell lines, or using other tumor-forming cancer cell lines, according to what type of cancer is of particular interest to the user. When dosed orally or intravenously, compounds induce rapid and complete tumor responses (CR) that are durable for several weeks after the end of treatment in all animals bearing H889 (SCLC) or RS4;11 (ALL) tumors. Similar treatment of mice bearing H146 SCLC tumors can induce rapid regressions in the animals.
9
Cell Line Culturing Techniques
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PC3 and LNCaP were purchased from Procell (Wuhan, China), DU145, RWPE-1, and human embryonic kidney 293E (HEK293E) cells from ATCC were gifted by Dr. Qi Li (The First Affiliated Hospital of Zhengzhou University, Zhengzhou, China) and cultured in a humidified environment at 37 °C under 5% CO2 using their respective media. RWPE-1 cells were maintained in keratinocyte SFM (1×) (Invitrogen, cat. 17,005,042). PC3 and LNCaP cells were cultured in RPMI 1640 supplemented with 15% fetal bovine serum (FBS). DU145 and HEK293E cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 15% FBS. LNCaP and PC3 cells stably overexpressing control and INMT vectors were cultured in RPMI 1640 supplemented with 15% FBS and hygromycin (150 µg/ml) [30 (link), 31 (link)]. LNCaP and PC3 cells stably expressing control shRNA (Sigma-Aldrich, cat. SHC016) and INMT shRNA (Sigma-Aldrich, cat. EHU138831) were cultured in RPMI 1640 supplemented with 15% FBS and puromycin (15 µg/ml) [31 (link)].
10
TMEFF2 Knockdown in LNCaP and 22Rv1 Cells
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The LNCaP and 22Rv1 cell lines were purchased from American Type Culture Collection (ATCC; Manassas, VA) and cultured as recommended. Dihydrotestosterone (DHT; Sigma, Burlington,MA) was used at a concentration of 10 nM. For TMEFF2 knockdown, LNCaP and 22Rv1 cells were transduced with pLKO.1 lentiviral vectors with antisense TMEFF2 sequences shTMEFF2–0 (TRCN0000073518), shTMEFF2–1 (TRCN0000073519) and shTMEFF2–2 (TRCN0000073521). See Additional file 1 : Table S7 for sequences.
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