The development of castration-resistant (CR) Myc-CaP cell line has been reported previously [26 (link)]. CR Myc-CaP cell line was cultured in DMEM (Mediatech, Inc.) with 10% FBS. The 5T4-tranfected murine B16-F10 melanoma cell line (B16-h5T4) [27 (link)] was kindly provided by Peter Stern (Paterson Institute for Cancer Research, Manchester, UK) and was cultured in R10 medium (RPMI-1640 with Ultra glutamine (BioWhittaker/Lonza, Wokingham, UK); supplemented with 10% fetal bovine serum (Fisher Scientific, Pittsburgh, PA), 1 mM sodium pyruvate, 10 mM HEPES, 0.1 mg/ml gentamicine sulfate and 50 μM β-mercaptoethanol). The CR Myc-CaP and B16-h5T4 cell lines were tested to be mycoplasma-free; no other authentication assay was performed.
Rpmi 1640 with ultraglutamine
Manufactured by Lonza
Sourced in Denmark
RPMI 1640 with UltraGlutamine is a cell culture medium designed to support the growth and maintenance of a variety of cell types. It contains essential amino acids, vitamins, and other nutrients necessary for cell proliferation. The inclusion of UltraGlutamine provides a stable source of L-glutamine, a key component for cellular metabolism.
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Lab products found in correlation
Streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin, by Thermo Fisher Scientific (2 mentions)
Newborn calf serum, by Sartorius (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
X vivo 20, by Lonza (1 mentions)
Lymphoprep, by Axis-Shield (1 mentions)
Interleukin 4 (il 4), by Immunotools (1 mentions)
Gm csf, by Immunotools (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Ethylene diamine tetraacetic acid (edta), by Thermo Fisher Scientific (1 mentions)
Biowhittaker, by Lonza (1 mentions)
Human serum, by Sartorius (1 mentions)
Wistar rats, by Taconic Biosciences (1 mentions)
Hct116, by American Type Culture Collection (1 mentions)
Medium m3 base, by INCELL (1 mentions)
Sw480, by American Type Culture Collection (1 mentions)
Sw620, by American Type Culture Collection (1 mentions)
Dmem 4.5 g l glucose, by Lonza (1 mentions)
Ncm460, by INCELL (1 mentions)
5 protocols using rpmi 1640 with ultraglutamine
1
Castration-Resistant Myc-CaP and B16-h5T4 Cell Lines
2
Generation and Maturation of Monocyte-Derived Dendritic Cells
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For generation of moDC, 20 to 45 ml heparinized blood were collected from each participant and handled within 2 hours. Peripheral blood mononuclear cells were isolated by discontinuous gradient centrifugation using Lymphoprep (Axis Shield PoC AS, Oslo, Norway) and monocytes isolated by plastic adherence in X-VIVO 20 (Lonza, Verviers, Belgium). Nonadherent cells were removed after 1 hour of incubation at 37°C and 5% carbon dioxide. Remaining monocytes were cultured in RP10 medium (RPMI 1640 with Ultraglutamine (Bio Whittaker, Lonza), 10% FCS Gold (PAA, Pasching, Austria), 1% penicillin–streptomycin (Gibco, Invitrogen Corporation, Paisley, UK)) supplemented with 100 ng/ml granulocyte–macrophage colony-stimulating factor (GM-CSF; ImmunoTools GmbH, Friesoythe, Germany) and 20 ng/ml IL-4 (ImmunoTools GmbH). Cytokines were replenished every 2 or 3 days. After 5 to 6 days in culture, maturation was induced in one part of the moDC by adding 1 μg/ml TLR7/8 ligand CL097 (InvivoGen, San Diego, CA, USA) for 48 hours.
3
Neonatal Rat Islet Isolation and Culture
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Neonatal rat islets of Langerhans were isolated from 4-day-old Wistar rat pups (Taconic, Lille Skensved, Denmark), as previously described [20] . All mice were housed according to the Principles of Laboratory Care. The isolated islets were pre-cultured for 7-10 days in RPMI 1640 with UltraGlutamine (Lonza, Vallensbaek, Denmark) and supplemented with 10% newborn calf serum (Biological Industries, Kibbutz Beit Haemek, Israel), 100 U/ml penicillin and 100 μg/ml streptomycin (Gibco, Life Technologies, Taastrup, Denmark) in 5% CO 2 at 37°C. For experiments, rat islets were cultured as intact and free-floating in medium supplemented with 2% human serum (BioWhittaker, Lonza) or as single cells following digestion with 0.2% trypsin (Gibco) and 10 mmol/l EDTA (Gibco) in Hanks' balanced salt solution (HBSS). Dispersed islets were cultured on coverslips coated with bovine corneal extracellular matrix (ECM; Biological Industries) in the medium described above containing 2% human serum.
4
Isolation and Transduction of Rat Pancreatic Islets
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Pancreatic islets from 8- to 10-week-old male Wistar rats (Taconic, Lille Skensved, Denmark) were isolated by collagenase digestion and hand-picked under a stereo microscope [13 (link)]. The isolated islets were precultured for 24 h before adenoviral transduction in RPMI 1640 with UltraGlutamine (Lonza, Vallensbaek, Denmark) supplemented with 10% newborn calf serum (Biological Industries, Kibbutz Beit Haemek, Israel), 100 U/ml penicillin and 100 μg/ml streptomycin (Life Technologies, Paisley, UK) in 5% CO2 at 37°C. All animal experiments were approved by the local ethics committee and performed in accordance with the Guide for the Care and Use of Laboratory Animals [14 ].
5
Culturing Colorectal Cancer Cell Lines
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The human colorectal carcinoma HCT116, colorectal adenocarcinoma HT29, HCT8, SW480, SW620, LoVo and SW48 cell lines were obtained from the American Type Culture Collection (ATCC). Some cell lines were grown in RPMI 1640 with ultraglutamine (Lonza, Levallois, France) (HCT116, HT29, HCT8, SW480) or in DMEM 4.5g/L glucose (Lonza) (SW620, LoVo, SW48) supplemented with 10% (vol/vol) fetal bovine serum (FBS; Lonza). The human normal mucosa cell line NCM460 was obtained from INCELL corporation LLC (San Antonio, TX) and was grown in M3Base medium (INCELL corporation LLC) supplemented with 10% FBS. All cell lines were grown in an atmosphere of 95% air and 5% CO2 at 37°C.
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