Vero E6 and BSC-40 cells (from ATCC) were maintained DMEM/F-12 medium with 10% fetal calf serum (FCS), 1X antibiotic-antimycotic, and 1X Glutamax (ThermoFisher). CHO-K1 cells (ATCC) were grown in F12K Medium (ATCC 30–2004) with 10% FCS. JUNV Candid #1 was received from Dr. Paessler (UTMB, Galveston, TX). Pathogenic JUNV (Romero) and MACV (Carvallo) were from Centers for Disease Control and Prevention (CDC, Atlanta, GA). MACV/Carvallo was passaged twice in Vero cells to prepare challenge stock at Texas Biomedical Research Institute (TBRI, San Antonio, TX). Titration of JUNV and MACV was performed by infecting Vero E6 monolayers under agarose overlay [42 (link)]. Inactivated (gamma-irradiated) MACV (Carvallo) and recombinant vaccinia virus encoding MACV GPC (rVACV-MACV GPC) [43 (link)] were received from BEI Resources.
Bsc 40 cells
Manufactured by American Type Culture Collection
Sourced in United States
The BSC-40 cells are a type of immortalized cell line derived from African green monkey kidney cells. They are commonly used in research applications as a model system for various cellular and molecular studies. The BSC-40 cells maintain a stable and consistent phenotype, making them a valuable tool for experiments that require a reliable and reproducible cell line.
Automatically generated - may contain errors
Lab products found in correlation
Vero e6 cell, by American Type Culture Collection (3 mentions)
Vero cell, by American Type Culture Collection (2 mentions)
Cpxv brighton red, by BEI Resources (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Cho k1 cells, by American Type Culture Collection (1 mentions)
F 12k medium, by American Type Culture Collection (1 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
Hepes, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Tnm fh medium, by Gemini Bio (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Nih3t3 cells, by American Type Culture Collection (1 mentions)
P815 mastocytoma cells, by American Type Culture Collection (1 mentions)
C myc clone 9e10, by Santa Cruz Biotechnology (1 mentions)
Actin clone 1 19, by Santa Cruz Biotechnology (1 mentions)
Celltiter 96 non radioactive cell proliferation assay mtt, by Promega (1 mentions)
Fetal bovine serum (fbs), by Corning (1 mentions)
Crizotinib, by Selleck Chemicals (1 mentions)
Temozolomide, by Merck Group (1 mentions)
10 protocols using bsc 40 cells
1
Protocols for Arenavirus Cell Culture
2
Viral Glycoprotein Expression Protocols
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BSC40 cells (ATCC CRL-2761) and Vero.E6 cells (ATCC CRL-1586) were grown in Dulbecco’s modified Eagle’s medium (DMEM; Life Technologies) which was supplemented with antibiotics (100 units/ml penicillin-100 µg/ml streptomycin [Pen-Strep]; Gibco), buffer solution (HEPES [1 M]; Gibco), and 10% fetal bovine serum (FBS; HyClone). Sf-9 cells were passaged in TNM-FH medium (Gemini Bioproducts) with antibiotics (Pen-Strep) and 10% FBS, while BTI-TN-5B1-4 cells were grown in serum-free SFX-insect medium (HyClone) supplemented with antibiotics (Pen-Strep). Replication-competent recombinant VSV (VSV-LASV) was provided by Heinrich Feldmann at Rocky Mountain Laboratories (RML), National Institute of Allergy and Infectious Diseases (NIAID), and has been described previously (17 (link)– (link)19 (link)). Recombinant vaccinia viruses expressing the glycoproteins of various arenaviruses were obtained from the Biodefense and Emerging Infections Research Resources Repository (BEI Resources). These include recombinant vaccinia viruses that express LASV Josiah GPC (NR-15493), LCMV Armstrong 53b GPC (NR-15497), MACV Carvallo strain GPC (NR-15501), GOTV INH-95551 GPC (NR-15486), WWAV AV 9310135 GPC (NR-15508), and JUNV XJ13 GPC (NR-15691) (16 (link)).
3
Cell Lines Used in Breast Cancer Research
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TUBO mouse mammary carcinoma cells were kindly provided by Dr. L. Landuzzi (University of Turin) [53 (link)]. Charles River Laboratories (Wilmington, MA) documented the cell line identity and freedom from adventitious agents. 4T1, MDA-MB-231, and MCF7 cells were purchased from the ATCC, while MTHJ cells [54 (link)] were provided by Dr. K. Mossman (McMaster University). Breast cancer cells were cultured in Dulbecco’s modified Eagle’s medium (DMEM) containing 2mM L-glutamine, 100U/mL anti-mycotic/antibiotic, non-essential amino acids and 1mM sodium pyruvate supplemented with 20% fetal bovine serum (FBS) for TUBO cells or 10% FBS for the remaining cells. BSC-40 cells were purchased from ATCC in 2005 and grown in minimal essential medium supplemented as above with 5% FetalGro bovine growth serum (RMBIO). All cells were passaged <20 weeks, and regularly mycoplasma tested.
4
Propagation and Titration of Orthopoxviruses
5
Propagation and Titration of Orthopoxviruses
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VACV Dryvax (NIH, Lot# 4008284), VACV Western Reserve (VACV-WR; ATCC VR-119) and CPXV Brighton Red (BEI Resources, NR-88) were propagated and titered in monolayer cultures of BSC-40 cells (ATCC CRL-2761). MPXV Zaire was propagated in BSC-40 cells and titered on Vero cells (ATCC CCL-81). Bangladesh 1974 Solaiman strain of VARV was propagated in monolayer cultures of Vero E6 cells (ATCC CRL-1586). VACV and CPXV were manipulated under BSL-2 conditions by vaccinated personnel. MPXV was manipulated under BSL-2 conditions with BSL-3 precautions by vaccinated personnel. All experiments with live VARV were reviewed and approved by the World Health Organization Advisory Committee on Variola Virus Research (WHO ACVVR). Experiments with VARV were conducted in accordance with WHO ACVVR guidelines and within a biosafety level 4 laboratory.
6
Vaccinia Virus Titering and Infection
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Vaccinia virus strain WR was a generous gift from Richard Condit (University of Florida, FL). For viral titering assays, cells were seeded in 24-well plates and grown overnight, followed by addition of 10,000 plaque-forming units (PFU)/well vaccinia virus. At 24 h after infection, resulting vaccinia virus was harvested by freeze-thaw lysis of infected cells. The resulting supernatant was serially 10-fold diluted in 24-well plates in DMEM containing 10% FBS and overlaid on BSC40 cells (ATCC) at 80% confluency. After 48 h, the medium was aspirated, and the cells were stained with 0.1% Crystal Violet in 20% ethanol, and then de-stained with 20% ethanol. Initial viral concentrations were determined by manually counting plaques. For proteomic, RNA-seq and western blotting analyses, vaccinia virus was added at an MOI of 5 and incubated for the indicated time before cells were harvested.
7
Recombinant Virus Production Protocols
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The recombinant LCMV strain Cl13 (rCl13) and Armstrong (rARM) expressing the surface glycoprotein of the LCMV strain WE (WE-GP) and the WE-GP N121K variants (rCl13∗, rARM∗) have been described (Sommerstein et al., 2015 (link)) and were engineered by standard procedures (Flatz et al., 2006 (link)). Recombinant Vaccinia virus expressing VSVG and vesicular stomatitis virus serotype Indiana (VSV) have been described (Charan et al., 1987 (link), Pinschewer et al., 1999 (link)). For virus production and titration BHK-21 cells (Clone 13, ECAAC), BSC40 cells (ATCC) and NIH 3T3 cells (ATCC) were used and were confirmed to be mycoplasma-negative. Owing to their origin from renowned international repositories they were not authenticated.
8
Cell Line Procurement and Validation
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BHK-21 cells, HEK293T cells were purchased from ECACC (Clone 13, Cat #85011433), P815 mastocytoma cells (TIB-64), NIH 3T3 and BSC40 cells from ATCC. FreeStyle 293-F suspension culture cells were purchased from Invitrogen/ThermoFisher. LCMV-GP-expressing BHK-21 cells (BHK-21-GP) and 293T-GP cells have previously been described.41 (link) All cell lines were regularly tested for mycoplasma and were negative. Owing to their origin from renowned international repositories and vendors they were not authenticated.
9
Cell Culture and Viability Assay Protocol
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BSC40 cells (Cat# CRL-2761) were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA). Murine embryonic fibroblasts (MEFs) were a kind gift from Dr. Carl Atkinson. LLC-A9F1 (A9F1) cells were a kind gift from Dr. Mark Rubenstein. All cells were cultured in DMEM supplemented with 10% Fetal Bovine Serum and 1x Penicillin-Streptomycin-L-Glutamine (Corning, Oneonta, NY, USA). Cell viability was measured using the CellTiter-96 Non-Radioactive Cell Proliferation (MTT) assay (Promega, Madison, WI, USA) according to the manufacturer’s recommendations. Antibodies used for this study were purchased from Santa Cruz Biotech (Santa Cruz, CA) and include: actin (clone I19) and c-Myc (clone 9E10).
10
Glioblastoma Cell Line Characterization
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BSC40 cells (Cat# CRL-2761) were purchased from American Type Culture Collection (Manassas, VA, USA). U118, GL261 and T9 cells were purchased from Mediatech (Herndon, VA, USA). All cells were cultured in DMEM (10% fetal bovine serum +1 × penicillin–streptomycin–l – glutamine Corning (Oneonta, NY, USA). Temozolomide was purchased from Sigma-Aldrich Co. (St Louis, MO, USA). Crizotinib was purchased from Selleckchem (Houston, TX, USA). Bevacizumab was purchased from Genentech, Inc. (South San Francisco, CA, USA). Western blot samples were prepared as described by Bartee et al.24 (link) Cell viability was measured using the CellTiter-96 Non-Radioactive Cell Proliferation Assay (Promega Corporation, Fitchburg, WI, USA), referred to as the MTT assay for the purposes of this paper, according to manufacturer’s recommendations. Caspase-3 activity was measured using the commercially available Colorimetric Caspase-3 Assay Kit (CASP3C, Sigma-Aldrich Co.) according to the manufacturer’s protocol. Antibodies used for this study were purchased from Santa Cruz Biotechnology Inc., Dallas, TX, USA and include GAPDH (clone G-9, sc-365062), Caspase-3 (clone 3C119, sc-70497), p-AKT (Clone 104A282, sc-52940).
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