Two human derived MB cell lines, DAOY and D283, were procured from American Type Culture Collection (ATCC, Manassas, VA). Both cell lines were grown in Eagle’s minimal essential media (EMEM) supplemented with 5% fetal bovine serum (FBS) from HyClone. The culture conditions for cell propagation and various assays listed below were maintained at 37°C with 5% CO2 in humidified incubator. Deidentified conceptual tissues were obtained from the Birth Defects Research Laboratory at the University of Washington (UW) in Seattle in full accordance with state and federal guidelines (UW Human Subjects Division Institutional Review Board (IRB) # STUDY00000380). The use of deidentified conceptual tissues for the preparation of human neural cell cultures was reviewed and approved by the North Texas Regional IRB (protocol # 2007–121). Primary human astrocytes of non-cancerous origin from three biological donors were isolated and cultivated as described previously (Gardner et al., 2006 (link)). The cells were cultured in Dulbecco’s modified eagle medium/nutrient mixture F-12 (DMEM/F12–1:1) supplemented with 10% FBS (Peak Serum, Fort Collins, CO), and 1% each of penicillin–streptomycin–neomycin (PSN) and amphotericin B, a fungicidal (Sigma).
Penicillin streptomycin neomycin
Manufactured by Merck Group
Sourced in United States
Penicillin-streptomycin-neomycin is a broad-spectrum antibiotic solution used in cell culture applications. It contains three antibiotics: penicillin, streptomycin, and neomycin, which work together to prevent bacterial contamination in cell cultures.
Automatically generated - may contain errors
Lab products found in correlation
Neomycin, by Merck Group (3 mentions)
Penicillin, by Merck Group (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Sk n sh, by Merck Group (2 mentions)
Sk n sh cell line, by American Type Culture Collection (2 mentions)
β actin, by Santa Cruz Biotechnology (2 mentions)
L glutamine, by Merck Group (2 mentions)
Streptomycin, by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Multiskan fc, by Thermo Fisher Scientific (2 mentions)
Stemxyme, by Worthington (2 mentions)
Dmem f12 media, by Caisson (2 mentions)
Essential and non essential amino acids, by Thermo Fisher Scientific (2 mentions)
Dnase 1, by Roche (2 mentions)
Amphotericin b, by Merck Group (1 mentions)
Crystal violet, by Merck Group (1 mentions)
Rpmi medium, by Merck Group (1 mentions)
Dulbecco s phosphate buffered saline, by Merck Group (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
20 protocols using penicillin streptomycin neomycin
1
Culturing Human Neural Cell Lines
2
Differentiation of SK-N-SH cells into neuronal cells
3
Meniscus Endothelial Cell Culture
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Human primary meniscus endothelial cells were obtained by fluorescence-activated cell sorting, as described above. After sorting, the cells were resuspended in an EC culture medium (StemCell, #08000) containing penicillin-streptomycin-neomycin (Sigma-Aldrich, #P4083) at a density of 2 × 104 cells/well. Subsequently, the cells were cultured overnight in 48-well plates. After adding apelin (100 nM, Sigma-Aldrich, #A6469) and/or ML221 (blocking apelin receptor, 100 nM, Sigma-Aldrich, #SML0919) for 8 h, the cells were harvested for indepth analysis.
4
Cell Proliferation and Cytotoxicity Assay
5
Didymin and Dox Encapsulation Protocol
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HP-β-CD and β-CD were provided by Aladdin Reagent Co., Ltd. (Shanghai, China). Didymin used in the study was obtained by our team (Figure S1 , batch number: 20180412, 95.37% of purity). The extraction and isolation procedures were introduced in the Supplementary information . DOX was purchased from Calbiochem (Calbiochem, China). DMSO, MTT and penicillin-streptomycin–neomycin solution were obtained from Sigma (Sigma-Aldrich, St. Louis. MO, USA). RPMI 1640 Medium, DMEM Medium, FBS, and 0.25% trypsin–EDTA were purchased from Gibco (Gibco-BRL, Carlsbad, CA, USA). All other chemicals and buffer solutions were of analytical grade.
6
Investigating Lung Cancer Cell Lines
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ERL, tivantinib, dasatinib, osimertinib, olmutinib, crizotinib, and cabozantinib were purchased from MedChemExpress LLC (Princeton, NJ, USA). CDODA-Me was kindly donated by Dr. Stephen Safe (Texas A & M University, College Station, TX, USA). RPMI 1640 medium, fetal bovine serum (FBS), crystal violet, hank balanced salt solution (HBSS), and penicillin-streptomycin-neomycin were purchased from Sigma Aldrich (St. Louis, MO, USA). Primary antibodies were purchased from Cell Signaling Technologies (Danvers, MA, USA), and β-actin was purchased from Santa Cruz Biotechnology, Inc. (Carlsbad, CA, USA). The non-small cell lung cancer cell line, HCC827, was purchased from American Type Culture Collection (ATCC) (Manassas, VA, USA); the HCC827R cell line (ERL-resistant) was provided by Dr. Arun Rishi (Wayne State University, Detroit, MI, USA).
7
Culturing Human Periodontal Ligament Fibroblasts and Osteoblasts
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Human periodontal ligament fibroblasts (HPdLF, Lonza) and human primary osteoblasts (HOB, PromoCell) were cultured in Dulbecco’s modified Eagle’s medium (DMEM; Invitrogen) containing 10% FBS (Life Technology), 1% L-glutamine (Thermo Fisher Scientific), 1% penicillin/streptomycin/neomycin (Sigma Aldrich), 1% L-ascorbic acid (Sigma Aldrich) and 20 µg/mL dexamethasone (Sigma Adlrich) at 37 °C, 5% CO2 and 95% humidity. The starvation medium contained reduced FBS concentration (1%). When reaching confluence, cells were passaged by application of Accutase® solution (Sigma Aldrich). For experiments, cells were used at passages four to six.
8
Culturing HeLa and NHDF Cell Lines
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HeLa, a human epithelioid cervix carcinoma cell line, was obtained from ATCC (Virginia, USA). Cells were cultured in Minimum Essential Medium (MEM#m2275, Sigma, Buchs, Switzerland) supplemented with 1% l -glutamine (Sigma-Aldrich, Buchs, Switzerland), 1% non-essential amino acids (NEAA, PAN Biotech, Aidenbach, Germany), 10% fetal calf serum (FCS, Sigma-Aldrich, Buchs, Switzerland), 1% Penicillin Streptomycin Neomycin (PSN, Sigma-Aldrich, Buchs, Switzerland) and 1 mM sodium pyruvate (Sigma-Aldrich, Buchs, Switzerland). Cells were sub-cultured every 2–3 days upon 80% confluence and cultured in a humidified incubator at 37 °C with 5% CO2 atmosphere (standard culture conditions).
NHDF, a non-cancerous human skin fibroblast cell line, was purchased from Sigma-Aldrich (Buchs, Switzerland). Cells were cultured in Dulbecco's Modifies Eagle Medium – high glucose (#RNBG3787, Sigma, Buchs, Switzerland) supplemented with 10% FCS, 1% PSN and 1%l -glutamine. Cells were sub-cultured once a week upon 80% confluence and medium was exchanged every 2–3 days. Cells were cultured under standard conditions.
NHDF, a non-cancerous human skin fibroblast cell line, was purchased from Sigma-Aldrich (Buchs, Switzerland). Cells were cultured in Dulbecco's Modifies Eagle Medium – high glucose (#RNBG3787, Sigma, Buchs, Switzerland) supplemented with 10% FCS, 1% PSN and 1%
9
Cell Line Culture and Novel HDACi
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Established human cell lines derived from DLBCL, MCL and SMZL were cultured according to the recommended conditions. All media were supplemented with fetal bovine serum (10%), Penicillin-Streptomycin-neomycin (~5,000 units penicillin, 5 mg streptomycin and 10 mg neomycin/mL, Sigma) and L-glutamine (1%). Two novel HDACi were used: ITF-A and ITF-B (Italfarmaco S.p.A., Milan, Italy).
10
Differentiating SK-N-SH Cells into Neuron-Like Cells
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Human SK-N-SH cell line was obtained from American Type Culture Collection (ATCC #HTB-11). Cells were cultured in Eagle’s Minimum Essential Medium (EMEM; ATCC). This media was supplemented with 1 % penicillin-streptomycin-neomycin (Sigma) and 10 % (v/v) fetal bovine serum (FBS). Retinoic acid (RA; Sigma) was used to induce SK-N-SH cells [12 (link)] to differentiate into more neuron-like cells [13 (link)] because this cell line is a mixture of different cell types. Approximately 15,000 cells were cultured in T-75 flask (Corning) supplemented with EMEM media for two days, and then Retinoic acid (10 μM) was added. Cells were treated with RA for two weeks and media was replaced every three-four days [12 (link)]. Cultures were monitored visually using light microscopy for morphological changes, and evaluated for neuronal cell markers (NeuN, PSD95 and NCAM) during the differentiation process [12 (link), 14 (link)].
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