The female cell line HEK293T were used for lentivirus production and ACE2-expressing HEK293T cells (a gift from A. Balazs, Ragon Institute) used for lentivirus infection were maintained in advanced DMEM (Sigma-Aldrich) supplemented with 10% FBS, 1X Penicillin-Streptomycin-L-Glutamine mixture (Gibco), 1X non-essential amino acids (Gibco), 1X sodium pyruvate (Gibco), and 1X HEPES buffer (Corning) (D10). The human female B cell lines 721.221 were generated previously by γ-radiation of 721 cells and do not express HLA A and B alleles (Shimizu and DeMars, 1989 (link)). These cell lines were maintained in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% (v/v) FBS (Sigma-Aldrich) and 1X Penicillin-Streptomycin-L-Glutamine mixture (Gibco). TAP-deficient mono-allelic HLA class I-expressing 721.221 cells were generated as described previously (please see companion manuscript) and maintained in 5ug/mL blasticidin (Invivogen), 0.5 ug/ml puromycin (Invivogen) and 1.5 mg/ml G418 (Invivogen).
Penicillin streptomycin l glutamine mixture
Manufactured by Thermo Fisher Scientific
Penicillin-Streptomycin-L-Glutamine mixture is a sterile liquid media supplement used in cell culture applications. It provides a combination of the antibiotics penicillin and streptomycin, along with the amino acid L-glutamine. This mixture is commonly used to prevent bacterial contamination and support cell growth in various in vitro cell culture systems.
Automatically generated - may contain errors
Lab products found in correlation
Sodium pyruvate, by Thermo Fisher Scientific (3 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (3 mentions)
Fetal bovine serum (fbs), by Merck Group (3 mentions)
Rpmi 1640 medium, by Merck Group (3 mentions)
Advanced dmem, by Merck Group (3 mentions)
Hepes buffer, by Corning (2 mentions)
Blasticidin, by InvivoGen (2 mentions)
Puromycin, by InvivoGen (2 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
3 protocols using penicillin streptomycin l glutamine mixture
1
Maintaining HEK293T and 721.221 Cell Lines
2
Cell Line Maintenance and Engineering
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The female cell line HEK293T were used for lentivirus production and ACE2-expressing HEK293T cells (a gift from A. Balazs, Ragon Institute) used for lentivirus infection were maintained in advanced DMEM (Sigma-Aldrich) supplemented with 10% FBS, 1X Penicillin-Streptomycin-L-Glutamine mixture (GIBCO), 1X non-essential amino acids (GIBCO), 1X sodium pyruvate (GIBCO), and 1X HEPES buffer (Corning) (D10). The human female B cell lines 721.221 were generated previously by γ -radiation of 721 cells and do not express HLA A and B alleles (Shimizu and DeMars, 1989 (link)). These cell lines were maintained in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% (v/v) FBS (Sigma-Aldrich) and 1X Penicillin-Streptomycin-L-Glutamine mixture (GIBCO). TAP-deficient mono-allelic HLA class I-expressing 721.221 cells were generated as described previously (please see companion manuscript) and maintained in 5ug/mL blasticidin (Invivogen), 0.5 ug/ml puromycin (Invivogen) and 1.5 mg/ml G418 (Invivogen).
3
Generation and Maintenance of Human B Cell Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
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The human female B cell line 721.221 were generated previously by γ-radiation of 721 cells and do not express HLA A and B alleles (Shimizu and DeMars, 1989 (link)). These cell lines were maintained in RPMI-1640 medium (Sigma-Aldrich) supplemented with 10% (v/v) FBS (Sigma-Aldrich) and 1X Penicillin-Streptomycin-L-glutamine mixture (GIBCO). HEK293T cells used for lentivirus production were maintained in advanced DMEM (Sigma-Aldrich) supplemented with 10% FBS, 2mM L-glutamine (GIBCO), 1X non-essential amino acids (GIBCO) and 1X sodium pyruvate (GIBCO).
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