Patients provided consent for the use of biospecimens for research as approved by the Clinical Trial Ethics Committee of Dushu Lake Hospital Affiliated of Soochow University. Within 30 min of surgical resection at Dushu Lake Hospital Affiliated of Soochow University, human bladder tumor tissues were submerged in Roswell Park Memorial Institute 1640 media (Gibco, America) supplemented with 10% FBS, 1% insulin-transferrin-selenium, 1% GlutaMAX (Gibco, America), and 1% penicillin-streptomycin (Gibco, America) and divided into twelve sections, followed by intratumoral injection with PBS and CA-NPs at the dose of 1 μg Ce6 for another incubation of 1 h. Then some of them were irradiated at the density of 0.15 W/cm2 for 5 min. After another 1 h incubation, half of these samples were cut into slices by freezing microtome for immunofluorescence staining of TUNEL assay. And the others were dissociated with 1 mg/ml collagenase II and 0.1 mg/ml DNase I in 1640 medium at 37°C for 45 min after irradiation. The single cells were harvested and counted, and 1 × 106 cells suspended in staining buffer were used for TUNEL staining measured by flow cytometry.
1640 media
Manufactured by Thermo Fisher Scientific
Sourced in United States
1640 media is a powdered cell culture medium designed for the growth and maintenance of a variety of mammalian cell types. It provides a balanced salt solution and essential nutrients required for cell proliferation. 1640 media is commonly used in cell biology research and can be supplemented with additional components as needed for specific applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (16 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (5 mentions)
Imr 5 75, by Leibniz Institute DSMZ (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions)
Mycoalert mycoplasma detection kit, by Lonza (3 mentions)
Penicillin, by Thermo Fisher Scientific (3 mentions)
Streptomycin, by Thermo Fisher Scientific (3 mentions)
Glutamax, by Thermo Fisher Scientific (3 mentions)
Mda mb 361, by American Type Culture Collection (2 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Merck Group (2 mentions)
Sk n fi, by Leibniz Institute DSMZ (2 mentions)
Lsm 780, by Zeiss (2 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (2 mentions)
4t1 cell, by American Type Culture Collection (2 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol, by Merck Group (2 mentions)
Fluostar optima, by BMG Labtech (2 mentions)
Aspc 1, by Thermo Fisher Scientific (1 mentions)
43 protocols using 1640 media
1
Photodynamic Therapy for Bladder Cancer
2
Gastric Cancer Cell Line Culture
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The gastric cancer cell lines GES, SGC-7901, HGC-27, Ncl-N87, and SNU-1 were obtained from Fuheng company, Shanghai, China, and were cultured in 1640 media (Gibco) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin/streptomycin (Millipore) in a humidified incubator with 5% CO2 at 37° C.
3
A549 Cell Culture for EGFR Overexpression
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The non-small cell lung cancer cell line (A549) was reported to overexpress EGFR37 (link). A549 cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 media (Gibco, USA) (pH 7.2±0.2) containing 10% fetal bovine serum (FBS), penicillin (50 UI/mL) and streptomycin (50 UI/mL) at 37 °C in a humidified atmosphere with 5% CO2.
4
Cell Culture Protocols for MDA-MB-361 and U2OS
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MDA-MB-361 (American Type Culture Collection (ATCC)) was cultivated as previously described43 (link). U2OS (ATCC) was grown under similar conditions except Roswell Park Memorial Institute (RPMI) 1640 media (GibcoTM cat # 21875-034) and FBS (10%) was utilized. The identity of all cell lines was established by microsatellite analysis (Forensik GmbH, Germany). All cultures were routinely checked for mycoplasma contamination using a MycoAlert mycoplasma detection kit (Lonza, cat. # LT07-218). In general, cells were grown to 80% confluency prior to removal from the dish using trypsin (0.25%)/EDTA (0.02%) for sub-culturing or harvesting.
5
Culturing Human Pancreatic Cancer Cell Lines
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The human PC cell lines PANC-1, BxPC-3 and ASPC-1 were purchased from Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). PANC-1 and ASPC-1 were maintained in DMEM media and BxPC-3 were cultivated in 1640 media (GIBCO, NY, USA). All media contained 10% heat-inactivated fetal bovine serum (FBS, GIBCO, USA), 100 U/mL penicillin and 100 μg/mL streptomycin (GIBCO, USA). All cell lines were incubated 37 °C in a 5% CO2 atmosphere.
6
Cell Culture and Protein Analysis
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A549 and H1299 cells were cultured in 1640 media (GIBCO) supplemented with 10% FBS at 37 °C in 5% CO2 humidified incubators. MG132 (cat. C2211) was purchased from Sigma-Aldrich (St. Louis, MO, USA). Thymidine (cat. 6060) was acquired from Millipore (Burlington, MA, USA). GeneMut siRNA transfection reagent (cat. SL100568) was purchased from SignaGen Laboratories (Rockville, MD, USA). LipoMax plasmid transfection reagent (cat. 17052012) was purchased from SUDGEN (Bellevue, WA, USA). M-PER lysis buffer (cat. 78501), Dynabeads (cat. 10004D) and Clean-blot™ IP detection reagent (HRP) (cat. 21230) were purchased from Thermo Fisher (Waltham, MA, USA). RIPA buffer (cat. P0013B) and Cell cycle and apoptosis analysis kit (cat. C1052) were purchased from Beyotime (Shanghai, China). The protease inhibitor cocktail (cat. B14012) and Cell Counting Kit-8 solution (CCK-8, cat. B34304) were purchased from Bimake (Houston, TX, USA). HRP-labeled goat anti-mouse (cat. GB23301) and goat anti-rabbit secondary antibody (cat. purGB23303) were purchased from Servicebio (Wuhan, China).
7
Culturing Human Choriocarcinoma Cells for EV Isolation
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The human choriocarcinoma
cell line (JAr) from the first trimester trophoblasts was acquired
from the American Type Culture Collection (ATCC, HTB-144, Teddington,
U.K.). JAr cells were cultured in a T75 flask in Roswell Park Memorial
Institute (RPMI) 1640 media (Gibco, Scotland) supplemented with 1%
penicillin/streptomycin (P/S, Gibco 15140122, Bleiswijk, Netherlands),
1%l -glutamine (Sigma, 59202C, St. Louis), and 10% fetal
bovine serum (FBS, Gibco, 10500064) at 37 °C under moist 5% CO2-rich conditions. At 80% confluence, the conditioned medium
was removed, and the cells were washed with 10 mL of nonsupplemented
RPMI 1640 media to remove traces of FBS. Nonsupplemented RPMI 1640
medium was replaced with fresh RPMI 1640 medium supplemented with
1% penicillin/streptomycin, 1%l -glutamine, and 10% EV-depleted
FBS. Cells were cultured for 24 h at 37 °C and 5% CO2. After incubation, the conditioned medium was collected for EV isolation.
We have submitted all relevant data of our experiments to the EV-TRACK
knowledgebase (EV-TRACK ID: EV190091).43 (link)
cell line (JAr) from the first trimester trophoblasts was acquired
from the American Type Culture Collection (ATCC, HTB-144, Teddington,
U.K.). JAr cells were cultured in a T75 flask in Roswell Park Memorial
Institute (RPMI) 1640 media (Gibco, Scotland) supplemented with 1%
penicillin/streptomycin (P/S, Gibco 15140122, Bleiswijk, Netherlands),
1%
bovine serum (FBS, Gibco, 10500064) at 37 °C under moist 5% CO2-rich conditions. At 80% confluence, the conditioned medium
was removed, and the cells were washed with 10 mL of nonsupplemented
RPMI 1640 media to remove traces of FBS. Nonsupplemented RPMI 1640
medium was replaced with fresh RPMI 1640 medium supplemented with
1% penicillin/streptomycin, 1%
FBS. Cells were cultured for 24 h at 37 °C and 5% CO2. After incubation, the conditioned medium was collected for EV isolation.
We have submitted all relevant data of our experiments to the EV-TRACK
knowledgebase (EV-TRACK ID: EV190091).43 (link)
8
Cell Line Cultivation and Maintenance
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B16 melanoma cells (American Type Culture Collection (ATCC), Manassas, VA, USA) and Chinese hamster ovary-K1 (CHO, ATCC) cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM, ThermoFisher, Carlsbad, CA, USA) containing 10% foetal bovine serum (FBS, Carpricorn Scientific, Ebsdorfergrund, Germany), 100 U/mL penicillin, and 100 µg/mL streptomycin (Gibco BRL, Dublin, Ireland). CT-26 BALB/c colon carcinoma cells (provided by Dr. Kwon, Ajou University) [28 (link)] were cultured in Roswell Park Memorial Institute (RPMI) 1640 media (GIBCO) containing 10% FBS, 100 U/mL penicillin, and 100 µg/mL streptomycin. HEK-293T cells (provided by Dr. Kwon, Ajou University) were cultured in FreeStyleTM 293 expression medium (Gibco).
9
Establishment of Cisplatin-Resistant Ovarian Cancer Cell Lines
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Side population (SP) and non-SP human ovarian cancer cell lines were generated in our lab.[2 ] Human ovarian cancer SKOV3 and A2780 cell lines were generated in our lab too. The stable cisplatin-resistant cell lines SKOV3/DDP and A2780/DDP were established from SKOV3 and A2780 cells, respectively, by continuous exposure of the cells to increasing concentrations of cisplatin and routine maintenance in 1640 media (Gibco, Carlsbad, CA) supplemented with 10% fetal bovine serum (Corning, New York, NY), 2 μmol/L l -glutamine at 37 °C in a humidified atmosphere containing 5% CO2.
10
Cultivation and Characterization of Neuroblastoma Cell Lines
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The human NB-1, IMR-5/75 and SH-SY5Y neuroblastoma cell lines were purchased from the Deutsche Sammlung von Mikroorganismen und Zellkulturen (Braunschweig, Germany) and cultivated in Roswell Park Memorial Institute (RPMI) 1640 media (Gibco Thermo Fischer Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (MilliporeSigma, Burlington, MA, USA) and 500 µg/mL geneticin (G418, MilliporeSigma, Burlington, MA, USA). The human SK-N-BE(2) neuroblastoma cell line was purchased from the American Type Culture Collection (Manassas, VA, USA), and cultivated in Dulbecco’s Modified Eagle Medium (DMEM, Thermo Fischer Scientific, Waltham, MA, USA) supplemented with 10% fetal bovine serum (MilliporeSigma, Burlington, MA, USA) and 500 µg/mL G418 (MilliporeSigma, Burlington, MA, USA). Cell line identity was confirmed using short tandem repeat DNA genotyping (Eurofins Scientific SE, Luxemburg, Luxemburg), and cultures were routinely tested for mycoplasma contamination using the PlasmoTest Kit (Thermo Fischer Scientific, Waltham, MA USA). All cells were cultured in a humidified atmosphere containing 5% CO2 at 37 °C.
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