THP1 cells were purchased from ATCC and cultured in RPMI-1640 media supplemented with l -Glutamine and sodium pyruvate (Sigma R8758) in addition to 10% fetal bovine serum and 1% penicillin/streptomycin in a 5% CO2 incubator. THP1 cells were differentiated using 100 ng/mL PMA (Sigma P1585) for 48 h. HEK293 and HEK293FT cells were obtained from ATCC and Thermo Fisher, respectively, and cultured using Dulbecco’s modified Eagle’s medium (DMEM) supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin in a 5% CO2 incubator. HEK293-RyR2 cells were provided by Dr. Wayne Chen47 (link). These cells possess doxycycline-inducible RyR2 expression, which enables spontaneous calcium oscillation in response to elevated extracellular calcium via store-overload induced calcium release47 (link),64 (link)–66 (link). Cells were cultured using DMEM supplemented with 10% fetal bovine serum and 1% penicillin/streptomycin in a 5% CO2 incubator. Treatment with doxycycline (1 µg/mL) for 24 h was used to induce RyR expression.
L glutamine and sodium pyruvate
Manufactured by Merck Group
Sourced in United States
L-Glutamine and sodium pyruvate are commonly used compounds in cell culture media. L-Glutamine is an amino acid that serves as an energy source for cells, while sodium pyruvate is a salt of pyruvic acid that can also be utilized as an energy source. These compounds are essential for maintaining the viability and growth of various cell lines in laboratory settings.
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Lab products found in correlation
Rpmi 1640 media, by Merck Group (1 mentions)
Hek293, by American Type Culture Collection (1 mentions)
Hek293ft, by American Type Culture Collection (1 mentions)
Thp 1 cell, by American Type Culture Collection (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Dmem high glucose, by Merck Group (1 mentions)
Penicillin streptomycin, by Euroclone (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Mcf 7, by Merck Group (1 mentions)
L glutamine penicillin streptomycin solution, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Bovine serum, by Thermo Fisher Scientific (1 mentions)
Dulbecco s modification of eagle s medium dmem, by Merck Group (1 mentions)
3 protocols using l glutamine and sodium pyruvate
1
Cell Culture Protocols for THP1, HEK293, and HEK293-RyR2
2
Cell Culture Protocol for MDA-MB-231 and MCF-7
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All the experiments were performed with MDA-MB-231 (ECACC 92020424) and MCF-7 (Sigma-Aldrich, St. Louis, MO, USA, ECACC, #92020424) cells. Cells were cultured in a Dulbecco’s modified Eagle medium (DMEM-High Glucose), with L-glutamine and sodium pyruvate (Sigma-Aldrich, St. Louis, MO, USA), and supplemented with 10% (v/v) of fetal bovine serum (FBS) (Sigma-Aldrich, St. Louis, MO, USA) and 1% (v/v) penicillin-streptomycin (EuroClone Spa, Pero (MI), Italy). Both cell lines and cells seeded on the substrates were cultured in the incubator at 37 °C, 5% CO2.
3
Antiviral Activity of 1,2,3-Triazole Derivatives
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The 1,2,3-triazole derivatives were diluted at the concentration of 50 mM in dimethylsulfoxide (DMSO) and stored at -20°C. We tested ACV as a reference antiviral drug to compare our data.
These compounds were evaluated in vitro on human fibroblast (HFL1 ATCC ® CCL-153 TM ) cells which were cultivated at 37°C and 5% CO 2, with Dulbecco's Modification of Eagle's medium (DMEM), with 4.5 g/l glucose, l-glutamine and sodium pyruvate (Sigma-Aldrich, Saint Louis, MO, USA), supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
To perform the plaque assay, we used rabbit skin (RS) cells cultivated at 37°C and 5% CO 2 with Minimum Essential Medium Eagle (MEM), with Earle's salts and l-glutamine (Sigma-Aldrich), supplemented with 5% bovine serum (Gibco) and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
The cells were cultivated until they achieved around 90% of confluence when they were transferred to plates of 24 or 96 wells depending on the assay.
HSV-1 strain 17syn+ was used for the experiments. We also tested the compounds on the ACV-resistant clinical strain HO-1 (kindly provided by the D Phelan, College of Medicine, University of Florida, personal communication). Virus stock cultures were prepared from supernatants of infected cells and stored at -80°C until use.
These compounds were evaluated in vitro on human fibroblast (HFL1 ATCC ® CCL-153 TM ) cells which were cultivated at 37°C and 5% CO 2, with Dulbecco's Modification of Eagle's medium (DMEM), with 4.5 g/l glucose, l-glutamine and sodium pyruvate (Sigma-Aldrich, Saint Louis, MO, USA), supplemented with 10% fetal bovine serum (Gibco, Carlsbad, CA, USA), and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
To perform the plaque assay, we used rabbit skin (RS) cells cultivated at 37°C and 5% CO 2 with Minimum Essential Medium Eagle (MEM), with Earle's salts and l-glutamine (Sigma-Aldrich), supplemented with 5% bovine serum (Gibco) and 1% of l-glutamine-penicillin-streptomycin solution (Sigma-Aldrich).
The cells were cultivated until they achieved around 90% of confluence when they were transferred to plates of 24 or 96 wells depending on the assay.
HSV-1 strain 17syn+ was used for the experiments. We also tested the compounds on the ACV-resistant clinical strain HO-1 (kindly provided by the D Phelan, College of Medicine, University of Florida, personal communication). Virus stock cultures were prepared from supernatants of infected cells and stored at -80°C until use.
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