Human NSCLC cell lines (A549, NCI-H460 and NCI-H1703) were purchased from the Korean Cell Line Bank (KCLB, Seoul, Korea) and maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and antibiotic-antimycotic solution (Life Technologies, Seoul, Korea). Human hepatocellular carcinoma cell line Huh7 was also purchased from the KCLB. Human bronchial epithelial cell line BEAS-2B was purchased from the American Type Culture Collection (ATCC, Manassas, VA) and maintained in DMEM medium supplemented with 10% FBS and antibiotic-antimycotic solution (Life Technologies). H9 hESCs were cultured on mouse embryonic fibroblast (MEF) feeder cells as described previously [6 (link), 14 (link)]. Human PBMCs were isolated by the Ficoll-Paque Plus method (GE Healthcare, Seoul, Korea). FO myeloma cells were maintained in DMEM medium supplemented with 10% FBS and antibiotic-antimycotic solution (Life Technologies)
Antibiotic antimycotic solution
Manufactured by Thermo Fisher Scientific
Sourced in United States, United Kingdom, Germany, Australia, Canada, Japan, Austria, China, India, France, Italy, Brazil, Switzerland, Argentina, New Zealand, Netherlands
Antibiotic-antimycotic solution is a sterile, liquid product that contains a combination of antibiotics and antimycotic agents. The solution is designed for use in cell culture applications to prevent bacterial and fungal contamination.
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Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (520 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (152 mentions)
L glutamine, by Thermo Fisher Scientific (91 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (81 mentions)
Glutamax, by Thermo Fisher Scientific (74 mentions)
Rpmi 1640, by Thermo Fisher Scientific (54 mentions)
Fetal bovine serum (fbs), by Merck Group (51 mentions)
Trypsin edta, by Thermo Fisher Scientific (50 mentions)
Dmem f12, by Thermo Fisher Scientific (41 mentions)
Penicillin, by Thermo Fisher Scientific (35 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (34 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (33 mentions)
Streptomycin, by Thermo Fisher Scientific (32 mentions)
Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (28 mentions)
Foetal bovine serum (fbs), by Thermo Fisher Scientific (25 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (24 mentions)
Dexamethasone (dex), by Merck Group (24 mentions)
Penicillin streptomycin solution, by Thermo Fisher Scientific (24 mentions)
Amphotericin b, by Thermo Fisher Scientific (22 mentions)
Trypsin, by Thermo Fisher Scientific (22 mentions)
1 261 protocols using antibiotic antimycotic solution
1
Culturing Human Cell Lines for Research
2
Diverse Cell Culture Protocols for Tissue Engineering
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BMSCs (ATCC)were cultured in MEMα medium supplemented with 10% fetal bovine serum (FBS; Gibco), 1% antibiotic‐anti‐mycotic solution (Gibco) and 5 ng/ml bFGF (Gibco). Osteogenic differentiation media comprised the basal culture medium supplemented with 50 μM L‐ascorbic acid (Sigma), 10 mM β‐sodium glycerophosphate (Sigma) and 10 nM dexamethasone (Sigma). MC3T3 cells (Chinese Academy of Medical Sciences, China) were cultured in MEMα medium supplemented with 10% FBS (Gibco), 1% antibiotic‐anti‐mycotic solution (Gibco). RAW264.7 cells (RAW‐OCs, Chinese Academy of Medical Sciences)were cultured in DMEM medium supplemented with 10% FBS (Gibco) and 1% antibiotic‐anti‐mycotic solution (Gibco). The generation of colonic organoids from hiPSCs (CELLAPYBIO) involved a multistep technique whereby hiPSCs were directed to form definitive endoderm, hindgut structures and ultimate colonic organoids. hiPSCs were grown on matrigel (BD)‐coated six‐well plates in mTsSR1 medium (STEM CELL Technologies). All cells were maintained at 37°C with 5% CO2.
3
Isolation and Culture of hBMSCs and mBMMs
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This study was approved by the Institutional Review Board (IRB) of Yonsei University College of Medicine (IRB No: 4-2017-0232) and informed consent was obtained from all participants for human bone marrow samples used in this study. Bone marrow aspirates were obtained from the posterior iliac crests of seven adult donors (55–65 years old) and isolated as previously described52 (link). hBMSCs were maintained in low-glucose Dulbecco’s modified Eagle’s medium (DMEM-LG; Gibco) supplemented with 10% FBS (Gibco) and 1% antibiotic/antimycotic solution (Gibco) and were used within five passages. RAW264.7 cells (Korean Cell Line Bank, Seoul, South Korea) were maintained in high-glucose DMEM (Gibco) supplemented with 10% FBS (Gibco) and 1% antibiotic/antimycotic solution (Gibco). mBMMs were cultured in α-minimum essential medium (Gibco) containing 10% FBS and 1% antibiotic-antimycotic solution. Polaprezinc (CAS 107667-60-7, Santa Cruz Biotechnology, Santa Cruz, CA, USA) was dissolved in 1 M hydrogen chloride and used at a final concentration of 50 μM. ZnSO4 was purchased from Sigma-Aldrich.
4
Isolation and Culture of Human Endometrial Stromal Cells
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HESCs were obtained at the time of hysterectomy for uterine fibroids from normally cycling premenopausal women. Patients were not undergoing hormonal treatment at the time of surgery. All the samples were collected during the proliferative phase of the cycle. HESCs were isolated and cultured as previously described30 (link)31 (link)32 (link). Briefly, endometrial samples were collected in Dulbecco’s modified Eagle’s medium (DMEM)/F-12 containing Antibiotic-Antimycotic solution (Invitrogen). After enzymatic digestion by DNase I (Sigma) and Collagenase I a (Sigma), the stromal cells were separated from epithelial cells and passed into culture. Proliferating HESCs were cultured in maintenance medium of DMEM/F-12 containing 10% dextran-coated charcoal-treated FBS and 1% antibiotic–antimycotic solution (Life technologies). Confluent monolayers of HESCs were treated with or without 0.5 mM 8-bromo-cyclic adenosine monophosphate (8-br-cAMP; Sigma) and 10−6 M medroxyprogesterone acetate (MPA; Sigma). All experiments were conducted before the third passage of the cultures.
5
Breast Cancer Cell Line Cultivation and Stimulation
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Human breast cancer cell lines (MCF-7, ZR-75-1, BT474, SK-BR-3, MDA-MB-453, MDA-MB-231), a mammary epitherial cell line (MCF-10A), colorectal cancer cell lines (HCT116, SW480), and epidermoid carcinoma cell line (A431) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). The KPL-3C cells were established, characterized, and kindly provided by Dr. Jun-ichi Kurebayashi (Kawasaki Medical School)38 (link). All of the cell lines were cultured under the respective depositor's recommendations. The MCF-7 cells were seeded in 48-well plates (2 × 104 cells mL−1), 24-well plates (1 × 105 cells mL−1), 6-well plates (3 × 105 cells 2 mL−1), or 10-cm dishes (2 × 106 cells 10 mL−1) in MEM (Life Technologies, Rockville, MD, USA) supplemented with 10% FBS (Nichirei Biosciences, Tokyo, Japan), 1% antibiotic/antimycotic solution (Life Technologies), 0.1 mM NEAA (Life Technologies), 1 mM sodium pyruvate (Life Technologies), and 10 μg mL−1 insulin (Sigma, St. Louis, MO, USA). The cells were maintained at 37°C with 5% CO2. The next day, the medium was changed to phenol red-free DMEM/F12 (Life Technologies), supplemented with FBS, antibiotic/antimycotic solution, NEAA, sodium pyruvate, and insulin. After 24 h, the cells were treated with 10 nM 17-oestradiol (E2, Sigma) ± xanthohumol or peptides (e.g., ERAP).
6
In Vitro Scratch Assay for Endothelial Cell Migration
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Confluent monolayers of human LECs or murine MS-1 cells grown in 24-well plates were cultured for 24 h in starvation medium. In the case of human LECs and HUVECs, starvation medium consisted of EBM-2 (Lonza) containing 2% FCS (Life Technologies) and 1% antibiotic-antimycotic solution (Life Technologies). In the case of primary dermal LECs and MS-1, starvation medium consisted of DMEM medium (Life Technologies) supplemented with 5% FCS (Life Technologies) and 1% antibiotic-antimycotic solution (Life Technologies). Subsequently, two cross-shaped scratches were made in each well. Monolayers were washed with PBS and 500 μl of starvation medium supplemented with αALCAM (R&D Systems), I/F8-Fc, isotype control, or KSF-Fc (all 10 μg/ml) in combination with VEGF-A (20 ng/ml; PeproTech) was added. Pictures were taken immediately after scratching and 12–24 h later using an Axiovert 200 M microscope and a 5 × objective (NA of 0.12) (Carl Zeiss, Inc.). Afterwards, the percentage of the surface area closed after 16–24 h was calculated using TScratch software (53 (link)).
7
Investigating AANG Effects on mTECs
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The mouse tubular epithelial cells (mTEC) were cultured in DMEM/F12 (Gibco, CA), supplemented with 10% FBS (Gibco, CA) and 1% antibiotic/antimycotic solution (Life Technologies, USA). The mouse mesangial cells (mMES) (MES13 ATCC, Manassas, VA, USA) was maintained in a 3:1 mixture of DMEM and Ham's F-12 medium containing 5% FBS, and 1% antibiotic/antimycotic solution (Life Technologies, Grand Island, NY, USA)10 (link), 12 (link), 15 (link). mTECs were seeded and treated with combination of AANG (20 µM AA+200 µM NG) in a 6-well plate for 6-hours, then stimulated with advanced glycation end-products (AGE) for 24-hours. Subsequently, stimulated mTECs were employed for western blot analysis.
8
Culturing Breast Cancer Cell Lines
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The human breast adenocarcinoma cell line MCF-7 and human breast carcinoma cell lines T-47D and MDA-MB-231 were purchased from the Leibniz Institute DSMZ-German Collection of Microorganisms and Cell Cultures (Braunschweig, Germany). MCF-7 was grown in 90% RPMI 1640 (Life technologies, Gibco®) supplemented with 15% fetal bovine serum (FBS, Life technologies, Gibco®), 1% Antibiotic-Antimycotic solution (AA, Life technologies, Gibco®), 1% MEM Non-Essential Amino Acids solution (Life technologies, Gibco®), 1 mM sodium pyruvate (Sigma-Aldrich, St. Louis, MO, USA) and 10 μg/mL human insulin (Sigma-Aldrich, St. Louis, MO, USA); T-47D cell line was grown in 85% RPMI 1640 supplemented with 15% fetal bovine serum (FBS), 1% Antibiotic-Antimycotic solution and 10 μg/mL human insulin, while the MDA-MB-231 cell line was grown in 85% RPMI 1640 supplemented with 15% fetal bovine serum (FBS) and 1% Antibiotic-Antimycotic solution. Human mammary epithelial cells (HMEC) were purchased from Life technologies (Gibco®) and grown in HUMEC serum-free medium supplemented with 20 μg/mL of Antibiotic-Antimycotic solution (Life technologies, Gibco®). All cells were incubated in a humidified atmosphere containing 5% CO2 and 95% air at 37 °C. Culture media was changed every 2 days and the cultures were passaged with 0.25% trypsin-EDTA (Life technologies, Gibco®) when 80% of confluence was achieved.
9
Glioblastoma cell culture protocol
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A172 and U251 cells were obtained from the European Collection of Cell Cultures (EACC, UK) in 2014 and authenticated in 2020 by Cell Bank Australia using short tandem profiling. Cells were cultured in DMEM supplemented with 10% FBS and Antibiotic-Antimycotic solution (Life Technologies, Carlsbad, CA, USA) at 37 °C and 5% CO2. Patient-derived HW1, RN1, MMK1 and RKI1 glioblastoma stem cells34 (link) were cultured in KnockOut DMEM/F-12 supplemented with StemPro NSC SFM, 2 mM GlutaMAX-ICTS, 20 ng/mL EGF, 10 ng/mL FGF-β and Antibiotic-Antimycotic solution (all Life Technologies) as adherent cells on flasks coated with MatriGel (Corning, NY, USA). The protocols were approved by the Human Ethics Committee of the Royal Brisbane & Women’s Hospital (RBWH 2004/161). Cell cultures were routinely tested for mycoplasma infection and culturing did not exceed 15 passages.
10
Standardized Preparation of C. cayetanensis Oocysts
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C. cayetanensis oocysts stored in 2.5% potassium dichromate were kindly provided by Dr. Alexandre da Silva, US-FDA in May 2018 for use in this study. To prepare dilutions for spiking, the oocysts were washed by centrifugation to remove potassium dichromate, enumerated by a haemocytometer and diluted to the required concentration (either 200, 10 or 5 oocysts/25 µL, as per the US-FDA validation study [9 (link)]) in 0.1% Alconox (Sigma-Aldrich, Oakville, ON, Canada) solution supplemented with 1× antibiotic–antimycotic solution (Gibco™, Waltham, MA, USA). The quantity of oocysts in the prepared spiking stocks was then verified by counting two 25-µL aliquots on a welled microscopy slide. Cryptosporidium parvum oocysts (calf-passaged, Iowa isolate) were purchased from Waterborne, Inc. (New Orleans, LA, USA) for use as the DNA extraction positive control. C. parvum oocysts were enumerated by the haemocytometer and stored at 4 °C in 1× antibiotic–antimycotic solution (Gibco).
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