Human prostate tumor cell lines, VCaP, LNCaP were purchased from American Type Culture Collection (ATCC) and LAPC4 kindly provided by Dr Charles L. Sawyers (UCLA, CA). VCaP and LNCaP cells were maintained in DMEM (ATCC), and LAPC4 cells in RPMI-1640 (ATCC) supplemented with 10% of fetal bovine serum (Invitrogen) in a humidified CO2 (5%) incubator at 37 °C. Cell culture and cell line treatment studies have been performed in two independent sets of experiments.
Lapc4
Manufactured by American Type Culture Collection
Sourced in United States
The LAPC4 is a piece of laboratory equipment designed for cell culture applications. It is a cell line developed by the American Type Culture Collection (ATCC) for research purposes. The LAPC4 cell line is derived from human prostate cancer cells and is a commonly used model for prostate cancer research. This product provides researchers with a standardized and well-characterized cell line to study various aspects of prostate cancer biology and disease progression.
Automatically generated - may contain errors
Lab products found in correlation
Lncap, by American Type Culture Collection (29 mentions)
Du145, by American Type Culture Collection (12 mentions)
Rwpe 1, by American Type Culture Collection (8 mentions)
Lapc4, by Thermo Fisher Scientific (5 mentions)
Lncap, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (4 mentions)
Hek293t, by American Type Culture Collection (4 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Nci h660, by American Type Culture Collection (3 mentions)
Pnt1a, by American Type Culture Collection (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Lipofectatmine2000, by Thermo Fisher Scientific (2 mentions)
Dulbecco s modified eagle s medium, by Corning (2 mentions)
Lncap, by Corning (2 mentions)
Iscove s modified dulbecco s medium, by Corning (2 mentions)
Rpmi 1640 medium, by Corning (2 mentions)
Lapc 4, by Corning (2 mentions)
R1881, by PerkinElmer (2 mentions)
Charcoal stripped serum, by Merck Group (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
34 protocols using lapc4
1
Culturing Prostate Tumor Cell Lines
2
Prostate Cancer Cell Line Cultivation and Characterization
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Human prostate cancer cells, LAPC4, VCaP, DU145, PC-3, and LNCaP cells were obtained from the ATCC (Manassas, VA), which also provided authentication of all cell lines. The cells were grown in ATCC-recommended medium supplemented with 10% fetal bovine serum, 100 U/ml penicillin and 100 μg/ml streptomycin. The authenticated human prostate carcinoma C4-2 was purchased from UroCor. Inc (Oklahoma City, OK) and grown in T medium containing 5% FBS. Cells from each of cell line were frozen at early passages and kept in liquid nitrogen providing a stock for cultured cells that were used within 6 month after thawing. Tetracycline responsive PC-3 cells were maintained in media recommended by ATCC for parental PC-3 cells supplemented with 10% Tet system-approved FBS from Clontech (Mountain View, CA). All culture media were purchased from Life Technologies (Carlsbad, CA); fetal bovine serum (FBS) was purchased from Sigma-Aldrich (St. Louis, MO). Doxycycline (Dox) was from Clontech. Collagen type I, fibronectin, and Matrigel were from Becton Dickinson Biosciences (Bedford, MA). CellTracker™ Green CMFDA was from Life Technologies. Bisindolylmaleimide I (BIM-I) was purchased from AdipoGen (San Diego, CA) and phorbol 12-myristate 13-acetate (PMA) was purchased from Sigma-Aldrich. AZD5363 and LY294002 were purchased from Selleckchem (Houston, TX).
3
Culturing and Maintaining Prostate Cancer Cell Lines
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Commercially available PCa cell lines (RWPE-1, LNCaP, 22Rv1, VCaP, LAPC4, PC3, DU145, NCI-H660, C4-2) were purchased from ATCC and maintained according to ATCC protocols. WCM154 and WCM155 CRPC-NE cell lines have been previously established and were maintained in two-dimensional monolayer culture according to the previously described protocol29 (link). LNCaP-AR cells were a kind gift from Dr. Sawyers and Dr. Mu (Memorial Sloan Kettering Cancer Center) and were cultured as previously described12 (link). MSKCC-PCa3 CRPC-Adeno cells were a kind gift from Dr. Chen (Memorial Sloan Kettering Cancer Center) and were maintained identically to WCM154 and WCM155 cells. All cell lines used and their phenotype are listed in Supplementary Table 4 . Cell cultures were regularly tested for Mycoplasma contamination and confirmed to be negative.
4
Comparison of Prostate Cell Lines
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A panel of immortalized human prostatic epithelial cell (BPH-1, RC-165N, PWR-1E), and prostate cancer cell lines (LNCaP and its metastatic C4 sublines, LAPC-4, VCaP, PC-3 and PC-3M, DU145, CA-HPV-10, ARCaPM) were used in this study. PWR-1E, LNCaP, LAPC-4, PC-3, and DU145 were obtained from ATCC (Manassas, VA), ARCaPM from Novicure Biotechnology (Birmingham, AL) whereas other cell lines were provided by the original investigators (BPH-1 from Dr S. Hayward; C4 sublines from Dr E.T. Keller; RC-165N from Dr J.S. Rhim; VCaP from Dr K. Pienta). PWR-1E, RC-165N and CA-HPV-10 were cultured in keratinocyte serum-free medium supplemented with bovine pituitary extract and recombinant epidermal growth factor, whereas other cell lines were maintained in different media as described previously [56 (link), 57 (link)].
5
Cell Culture and Transfection Protocols
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LNCaP, PC-3, LAPC-4 and 293T cells were purchased from ATCC (Manassas, VA). LNCaP, PC-3 cells were cultured in RPMI 1640 medium (Corning cellgro) supplemented with 10% fetal bovine serum (FBS) (Thermo Fisher Scientific). LAPC-4 cells were cultured in Iscove's modified Dulbecco's medium (Corning cellgro) supplemented with 15% FBS (Thermo Fisher Scientific). 293T cells for lentiviral packaging were cultured in Dulbecco's modified Eagle's medium (Corning cellgro) supplemented with 10% FBS (Thermo Fisher Scientific). Transfections were performed using Lipofectatmine2000 (Thermo Fisher Scientific), following manufacturer's instructions. 75-90% transfection efficiencies were achieved.
6
Cell Line Characterization and Transfection
7
Cell Line Maintenance and Handling
8
Prostate Cancer Cell Lines Cultivation
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PC3, DU145, LAPC4, LNCaP and RWPE‐1 cell lines were purchased from ATCC. The cell lines have been recently authenticated by short tandem repeat (STR) profiling at the Shanghai Biowing Applied Biotechnology Company. PC3 and DU145 cells were cultured in Dulbecco's modified Eagle's medium (DMEM; Gibco) containing 10% foetal bovine serum (FBS), 100 U/mL penicillin and 100 μg/mL streptomycin. LAPC4 and LNCaP cells were cultured in RPMI‐1640 medium (Gibco) supplemented with 10% FBS, 100 U/mL penicillin and 100 μg/mL streptomycin. RWPE‐1 cells were cultured in keratinocyte serum‐free medium (K‐SFM, Gibco) supplemented with growth supplement provided with the K‐SFM kit, 100 U/mL penicillin and 100 μg/mL streptomycin. The culture medium was refreshed every 2 days. In experimental cultures, cells were treated with indicated concentrations of pentamidine (Selleck, S4007) or 1.5 mmol/L N‐acetyl‐l ‐cysteine (NAC; Sigma, A9165). Sterile distilled water was served as the vehicle for pentamidine, and dimethyl sulfoxide (Sigma, D2650) was used as the solvent for NAC. NAC was administered with pentamidine at the same time. Other chemical reagents used in this study are listed in Table S1 .
9
Prostate Cancer Cell Line Modeling
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Androgen-dependent prostate cancer cell lines LNCaP and LAPC4 were purchased from ATCC. Enzalutamide (MDV3100) and JQ1 were purchased from Sigma-Aldrich.
Cells were cultured in RPMI or DMEM media with 5% charcoal-stripped FBS (CSS) and 1% penicillin streptomycin. Synthetic androgen R1881 was added at final concentration of 1 nM as androgen supplements. Four conditions were routinely used: (1) Basal (castration or ADT) condition, media with 5% charcoal-striped serum (CSS), (2) Hypoxia, 1% oxygen, (3) Androgen, CSS + 1 nM R1881, and (4) Combination (combo), hypoxia + androgen. In the anti-androgen therapeutic experiments, cells were cultured in the androgen condition (CSS + 1 nM R1881), and were treated by anti-androgen/AR drug enzalutamide (Enza) at the final concentration of 10 µM. The hypoxia or 1% oxygen condition was created in the cell culture incubator by replacing oxygen with compressed nitrogen. Sodium Bicarbonate (30 mM) was used to neutralize the hypoxia-induced lactate acid for experiments without involving metabolic measurement.
Cells were cultured in RPMI or DMEM media with 5% charcoal-stripped FBS (CSS) and 1% penicillin streptomycin. Synthetic androgen R1881 was added at final concentration of 1 nM as androgen supplements. Four conditions were routinely used: (1) Basal (castration or ADT) condition, media with 5% charcoal-striped serum (CSS), (2) Hypoxia, 1% oxygen, (3) Androgen, CSS + 1 nM R1881, and (4) Combination (combo), hypoxia + androgen. In the anti-androgen therapeutic experiments, cells were cultured in the androgen condition (CSS + 1 nM R1881), and were treated by anti-androgen/AR drug enzalutamide (Enza) at the final concentration of 10 µM. The hypoxia or 1% oxygen condition was created in the cell culture incubator by replacing oxygen with compressed nitrogen. Sodium Bicarbonate (30 mM) was used to neutralize the hypoxia-induced lactate acid for experiments without involving metabolic measurement.
10
Characterization of Prostate Cancer Cell Lines
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PC3, PC‐3M‐Luc, Du145, LNCaP, VCaP, LAPC4, 22Rv1 and the murine TRAMP‐C1 cell lines were obtained from ATCC (LGC Standards, Heidelberg, Germany) and maintained as previously described.15 Human cell lines were routinely authenticated by STR profiling as described before.16 Hamster BHK‐21 cells and murine L929 cells were obtained from ATCC and used for virus production and titration as described previously.17 BHK‐21 cells were maintained in Glasgow minimum essential medium (GMEM) (Gibco, Carlsbad, CA) supplemented with 10% heat inactivated FCS (PAA Laboratories, Vienna, Austria), 5% tryptose phosphate broth (Gibco), 100 units/ml penicillin (Gibco) and 0.1 mg/ml streptomycin (Gibco). L929 cells were maintained in high‐glucose DMEM (Lonza, Basel, Switzerland) supplemented with 10% heat inactivated FCS, 4 mM l ‐glutamine (Gibco), 100 units/ml penicillin and 0.1 mg/ml streptomycin.
VSV‐GP, VSVncp‐ΔG‐GFP*GP, VSV‐GP‐GFP and VSV‐GP‐Luc have been described before.17 A schematic representation of the VSV‐GP variants used is shown in Supporting Information 1.
VSV‐GP, VSVncp‐ΔG‐GFP*GP, VSV‐GP‐GFP and VSV‐GP‐Luc have been described before.
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