The human prostate cancer cell line LNCaP (DSMZ German Collection of Microorganisms and Cell Cultures GmbH, Braunschweig, Germany) was used for the establishment of tumor xenograft models. Cells were routinely screened for mycoplasma and authenticity was confirmed by short tandem repeat analysis. The cells were maintained as monolayer cultures in RPMI (Gibco, Carlsbad, US) containing 10% fetal bovine serum and Penicillin–Streptomycin (Gibco, Carlsbad, US) at 37 °C in a humidified CO2 atmosphere (5%).
Lncap
Manufactured by Leibniz Institute DSMZ
Sourced in Germany
The LNCaP cell line is a well-established prostate cancer cell line derived from a metastatic lesion of a human prostate carcinoma. It is widely used in cancer research as a model system for studying prostate cancer biology and drug development.
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Lab products found in correlation
Du145, by Leibniz Institute DSMZ (9 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (6 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (3 mentions)
Lncap, by Thermo Fisher Scientific (3 mentions)
Glutamax, by Thermo Fisher Scientific (3 mentions)
Du145, by Thermo Fisher Scientific (3 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
Mcf 7, by Leibniz Institute DSMZ (2 mentions)
Sk br 3, by Leibniz Institute DSMZ (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (2 mentions)
Hct116, by Leibniz Institute DSMZ (2 mentions)
Trypsin edta, by Thermo Fisher Scientific (2 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (2 mentions)
Hepes buffer, by Merck Group (2 mentions)
Iscove basal medium, by Harvard Bioscience (2 mentions)
Lncap, by Harvard Bioscience (2 mentions)
Hcc15, by Leibniz Institute DSMZ (1 mentions)
Ntera 2, by Leibniz Institute DSMZ (1 mentions)
Fetal bovine serum (fbs), by Leibniz Institute DSMZ (1 mentions)
19 protocols using lncap
1
Establishment of LNCaP Prostate Cancer Xenografts
2
Cell Culture Protocol for Cancer Lines
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PC-3, HCC15, NTERA-2, MCF-7, and LNCaP cells were purchased from DSMZ-German Collection of Microorganisms and Cell Cultures GmbH. The human TC cell line 1889c was kindly provided by Ehemann et al. [16 (link)]. Cells lines were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, and 100 U/mL penicillin/streptomycin (Gibco, Waltham, MA, USA) at 37 °C in a 5% CO2 humidified environment.
3
Cell Culture of Cancer Cell Lines
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The DU-145, LNCaP, T47D and SKBR3 cell lines were purchased from DSMZ (Braunschweig, Germany), and were cultured in RPMI supplemented with 10% fetal bovine serum (FBS), at 37 °C, 5% CO2. All media were purchased from Invitrogen (Carlsbad, USA) and all chemicals from Sigma (St. Louis, MO), unless otherwise stated.
4
Culturing Prostate Tumor and Endothelial Cells
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Human prostate tumour cell lines PC-3, DU-145 and LNCaP were obtained from DSMZ (Braunschweig, Germany). Tumour cells were grown and subcultured in RPMI 1640 (Gibco/Invitrogen; Karlsruhe, Germany). The medium contained 10% foetal calf serum (FCS), 2% 2-(4-(2-Hydroxyethyl)-1-piperazinyl)-ethansulfonsäure HEPES-buffer (1 M, pH 7.4), 2% glutamine and 1% penicillin/streptomycin. Subcultures from passages 7–11 were selected for experimental use.
Human endothelial cells (HUVEC) were isolated from human umbilical veins and harvested by enzymatic treatment with chymotrypsin. HUVEC were grown in Medium 199 (M199; Biozol, Munich, Germany), supplemented with 10% FCS, 10% pooled human serum, 20 μg/ml endothelial cell growth factor (Boehringer, Mannheim, Germany), 0.1% heparin, 100 ng/ml gentamycin and 20 mM HEPES-buffer (pH 7.4). Subcultures from passages 2–6 were selected for experimental use.
Human endothelial cells (HUVEC) were isolated from human umbilical veins and harvested by enzymatic treatment with chymotrypsin. HUVEC were grown in Medium 199 (M199; Biozol, Munich, Germany), supplemented with 10% FCS, 10% pooled human serum, 20 μg/ml endothelial cell growth factor (Boehringer, Mannheim, Germany), 0.1% heparin, 100 ng/ml gentamycin and 20 mM HEPES-buffer (pH 7.4). Subcultures from passages 2–6 were selected for experimental use.
5
Prostate Cancer Cell Lines in Angiotensin Signaling
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We used one slow growth, androgen-sensitive prostate cancer cell line LNCaP (DSMZ, Braunschweig, Germany) and invasiveness, androgen-insensitive cell line PC3 (ECACC). In 2014, the lines were authenticated by short-tandem repeat (STR) DNA profiling (LGC Standards Cell Line Authentication Service, Germany). Human prostate cancer cells were maintained as a traditional monolayer culture in RPMI (Gibco, Thermo Fisher Scientific Inc, Waltham, MA, USA) with 10% heat-inactivated fetal bovine serum (FBS) and standard supplements, such as: sodium pyruvate, L-glutamine, HEPES buffer and antibiotics (PSN). Ang-(1-9) (no. H-5038) and Ang-(3-7) (no. H-6965) were purchased from Bachem (Bubendorf, Switzerland), while angiotensin receptor inhibitors were purchased from TOCRIS (Bristol, UK): losartan (AT1 antagonist, no. 3798), PD123319 (AT2 antagonist, no. 1361), A779 (AT1-7/MAS antagonist, no. 5937), HIF142 (AT4/IRAP antagonist, no. 5627). The final concentrations of both angiotensins were 1nM whereas antagonists were used at 1000 nM. Given that these peptides are relatively quickly degraded, the experimental medium was changed every 24 h.
6
Prostate Cancer Cell Lines Cultivation
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Human, castration-resistant prostate tumor cell lines PC3, DU-145, and castration-sensitive LNCaP cells were purchased from DSMZ (Braunschweig, Germany). The prostate epithelial cell lines BPH-1 (from benign prostate hyperplasia) and PNT-1 (from normal prostate epithelium) were obtained from DSMZ and ATCC (Manassas, VA, USA). All cells were grown and subcultured in RPMI 1640 medium (Gibco/Invitrogen, Karlsruhe, Germany) augmented with 10% fetal calf serum (FCS), 2% HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid) buffer (1 M, pH 7.4), 2% glutamine, 1% penicillin/streptomycin at 37 °C in a humidified, 5% CO2 incubator.
7
Prostate Cancer Cell Line Characterization
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In the present study, four prostate cancer cell lines were used. The androgen-sensitive LNCaP (DSMZ GmbH, Braunschweig, Germany) and VCaP (ATCC, Manassas, VA, USA) cells are representative of prostate adenocarcinoma and the androgen-insensitive PC-3 (ATCC, Manassas, VA, USA) and CTM-LNCaP (subline of LNCaP) cells. To establish CTM-LNCaP, the parental LNCaP cells were cultivated for at least 3 months in phenol red-free RPMI medium supplemented with hormone-free charcoal-treated FBS.
All cells were cultured in RPMI-1640 antibiotic-free medium (Invitrogen, Waltham, MA, USA), supplemented with L-Glutamine, and grown in a 5% CO2 atmosphere at 37 °C. PC-3 and VCaP cells were cultured with 10% fetal bovine serum (FBS), CTM-LNCaP with 10% charcoal-treated fetal bovine serum (FBS), and LNCaP cells with 20% FBS.
All cells were cultured in RPMI-1640 antibiotic-free medium (Invitrogen, Waltham, MA, USA), supplemented with L-Glutamine, and grown in a 5% CO2 atmosphere at 37 °C. PC-3 and VCaP cells were cultured with 10% fetal bovine serum (FBS), CTM-LNCaP with 10% charcoal-treated fetal bovine serum (FBS), and LNCaP cells with 20% FBS.
8
Temsirolimus-resistant Prostate Cancer Cell Lines
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The human prostate tumor cell lines PC3, DU-145 (both castration-resistant), and LNCaP (castration-sensitive) were obtained from DSMZ (Braunschweig, Germany). Tumor cells were grown and subcultured in RPMI 1640 medium (Gibco/Invitrogen, Karlsruhe, Germany) supplemented with 10% fetal calf serum (FCS), 2% HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid) buffer (1 M, pH 7.4), 2% glutamine, 1% penicillin/streptomycin at 37 °C in a humidified, 5% CO2 incubator. The temsirolimus-resistant sublines developed over 12 months by continuous exposure to temsirolimus (LC Laboratories, Woburn, MA, USA), starting at 1 nmol/mL and increasing stepwise to 1 µmol/mL (PC3res, DU-145res, LNCaPres). The control cells remained untreated (PC3par, DU-145par, LNCaPpar).
9
Cell Culture Conditions for Cancer Cell Lines
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The human breast cancer cell lines MCF-7, SK-BR-3 and MBA-MB-231 (triple-negative), LNCap (human prostate carcinoma lymph node metastasis), HeLa (cervix carcinoma), HepG2 (hepatocellular carcinoma), and HCT116 (human colon carcinoma) were purchased commercially from DSMZ (Braunschweig, Germany). If not otherwise stated, all cell culture media and supplements were obtained from Life Technologies (Darmstadt, Germany).
Cells were cultured as described previously [36 (link)]. In detail, MCF-7 in RPMI medium containing 10% fetal bovine serum (FBS), 1 mM sodium pyruvate, and 0.1 M nonessential amino acids (NEAA); HeLa and HepG2 in RPMI medium containing 10% FBS and 2 mM Glutamax; MDA-MB-231 in Leibovitz’s L15 with 10% FBS; HCT116 and SK-BR-3 in McCoy’s 5A medium containing 10% FBS and 2 mM Glutamax; and LNCap in RPMI medium containing 10% FBS, 1 mM sodium pyruvate, 0.1 M NEAA, 2 mM Glutamax, and 0.01 M HEPES. All cell culture media were used without antibiotics. MCF-7, HCT116, SK-BR-3, LNCap, HeLa, and HepG2 were maintained at 37 °C in a humidified incubator with a 5% CO2 atmosphere and MDA-MB-231 cells were handled without gaseous exchange. The absence of mycoplasma in the cell culture was regularly tested using the Venor GeM qOneStep-Kit (Minerva-biolabs, Berlin, Germany).
Cells were cultured as described previously [36 (link)]. In detail, MCF-7 in RPMI medium containing 10% fetal bovine serum (FBS), 1 mM sodium pyruvate, and 0.1 M nonessential amino acids (NEAA); HeLa and HepG2 in RPMI medium containing 10% FBS and 2 mM Glutamax; MDA-MB-231 in Leibovitz’s L15 with 10% FBS; HCT116 and SK-BR-3 in McCoy’s 5A medium containing 10% FBS and 2 mM Glutamax; and LNCap in RPMI medium containing 10% FBS, 1 mM sodium pyruvate, 0.1 M NEAA, 2 mM Glutamax, and 0.01 M HEPES. All cell culture media were used without antibiotics. MCF-7, HCT116, SK-BR-3, LNCap, HeLa, and HepG2 were maintained at 37 °C in a humidified incubator with a 5% CO2 atmosphere and MDA-MB-231 cells were handled without gaseous exchange. The absence of mycoplasma in the cell culture was regularly tested using the Venor GeM qOneStep-Kit (Minerva-biolabs, Berlin, Germany).
10
Cancer Cell Lines and Nanoparticle Characterization
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The human cancer cell lines Detroit 562 (#CCL-138, pharyngeal carcinoma), FaDu (#HTB-43, squamous cell pharyngeal carcinoma), HT1080 (#CCL-121, fibrosarcoma), MIA PaCa-2 (#CRL-1420, pancreas carcinoma), NCI-H460 (#HTB-177, large cell lung carcinoma) and T98G (#CRL-1690, glioblastoma) were purchased from ATCC. The human cancer cell lines CAL-33 (#ACC 447, tongue squamous cell carcinoma), DU-145 (#ACC 261, prostate carcinoma), HEP-3B (#ACC 93, hepatocellular carcinoma), LNCaP (#ACC 256, prostate carcinoma) and PC-3 (#ACC 465, prostate adenocarcinoma) were purchased from DSMZ. The human cancer cell lines MDA-MB-231-luc-D3-H2LN (breast adenocarcinoma) and NCI-H460-Luc2 (large cell lung carcinoma) were purchased from Caliper. Cells were cultivated according to manufacturer’s recommendations and were controlled for mycoplasma by an independent laboratory (Clean Cells). NBTXR3 (Nanobiotix) is a sterile aqueous suspension of functionalized crystal hafnium oxide (HfO2) spherical nanoparticles with a size centered on 50nm, determined by dynamic light scattering technique (Zetasizer NanoZS, Malvern Instruments Ltd, Worcestershire, UK), bearing a marked negative surface charge (–50mV) in aqueous solution at pH 6–8,2 (link),4 (link) estimated by zeta potential analysis (Zetasizer NanoZS). Cisplatin (CDDP) was obtained from Sigma Aldrich.
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