Cell lines were cultured according to standard aseptic mammalian tissue culture protocols in 5% CO2 at 37 °C with regular testing for mycoplasma contamination using the MycoAlert Mycoplasma Detection kit (Lonza). HCT116, HEK293, HT1080, MCF7 and U2OS cells were cultured in DMEM supplemented with 10% fetal bovine serum (FBS) (Wisent) and 100 U ml−1 penicillin and 100 μg ml−1 streptomycin (Wisent). OP9 bone marrow stromal cells, before proliferation assays, were cultured in α-MEM medium with GlutaMAX (Invitrogen) containing 20% FBS (Wisent), 55 μmol liter−1 β-mercaptoethanol (Invitrogen) and 100 U ml−1 penicillin and 100 μg ml−1 streptomycin (Wisent). KMS-11 parental and TKO2 cells, a kind gift from J. Licht, University of Florida, were cultured in RPMI (Wisent) supplemented with 10% FBS (Wisent) and 100 U ml−1 penicillin and 100 μg ml−1 streptomycin (Wisent).
Streptomycin
Manufactured by Wisent
Sourced in Canada, United States, China
Streptomycin is a broad-spectrum antibiotic commonly used in laboratory settings. It is a product of the bacterium Streptomyces griseus and is effective against a variety of gram-positive and gram-negative bacteria. Streptomycin's core function is to inhibit protein synthesis in bacterial cells, thereby disrupting their growth and reproduction.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Wisent (124 mentions)
Fetal bovine serum (fbs), by Wisent (47 mentions)
Dulbecco modified eagle medium (dmem), by Wisent (23 mentions)
L glutamine, by Wisent (19 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (19 mentions)
Rpmi 1640, by Wisent (14 mentions)
Glutamine, by Wisent (7 mentions)
Streptomycin, by Thermo Fisher Scientific (6 mentions)
Hydrocortisone, by Thermo Fisher Scientific (6 mentions)
Penicillin, by Thermo Fisher Scientific (5 mentions)
Penicillin streptomycin, by Wisent (5 mentions)
Sodium pyruvate, by Wisent (5 mentions)
Epidermal growth factor (egf), by Thermo Fisher Scientific (5 mentions)
Basic fibroblast growth factor (bfgf), by Thermo Fisher Scientific (5 mentions)
Roswell park memorial institute (rpmi), by Wisent (4 mentions)
Rpmi 1640, by Thermo Fisher Scientific (4 mentions)
Bovine calf serum (bcs), by GE Healthcare (4 mentions)
L glutamate, by Wisent (4 mentions)
Phosphate buffered saline (pbs), by Wisent (4 mentions)
Amphotericin b, by Wisent (4 mentions)
137 protocols using streptomycin
1
Mammalian Cell Culture Protocols
2
Cell Culture of Thyroid and Kidney Cell Lines
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Cell lines 8505C (DSMZ, ACC‐219), KHM‐5 M (Expasy, CVCL_2975), and CAL62 (DSMZ, ACC‐448) were cultured with 10% fetal bovine serum (30044333, Gibco, USA), 100 units mL−1 penicillin and 100 µg mL−1 streptomycin (450‐201‐CL, WISENT, Canada) in RPMI‐1640 (11879020, Gibco, USA) medium. Cell line 293T (ATCC, CRL‐3216) was incubated with 10% fetal bovine serum (30044333, Gibco, USA), 100 units mL−1 penicillin and 100 µg mL−1 streptomycin (450‐201‐CL, WISENT, Canada) in DMEM (A1896701, Gibco, USA).
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Culturing HEK-293 and U2OS cells
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HEK-293 were cultured in DMEM high-glucose (GE Healthcare cat# SH30081.01) containing 10% bovine calf serum (GE Healthcare cat# SH30072.03), 2 mM L-glutamate (Wisent cat# 609065, 100 IU penicillin and 100 µg/ml streptomycin (Wisent cat# 450201). U2OS were cultured in DMEM high-glucose containing 10% tetracyclin-free fetal bovine serum (FBS) (Wisent cat# 081150) 2 mM L-glutamate, 100 IU penicillin and 100 g/ml streptomycin. Tetracyclin-free FBS was used to limit Cas9 expression. U2OS were starved for 2 hr in Earle’s balanced salts medium (Sigma cat# E2888).
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Maintenance of Cell Lines for Research
5
Cell Culture Protocols for Cancer Cell Lines
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HEK293FT (RRID:CVCL_6911 ), MDA-MB-231 (RRID:CVCL_0062 ), MDA-MB-468 (RRID:CVCL_0419 ), MDA-MB-436 (RRID:CVCL_0623 ) and COV362 (RRID:CVCL_2420 ), cells were cultured in high-glucose Dulbecco's Modified Eagle Medium (DMEM) containing HEPES buffer, penicillin, and streptomycin (Wisent cat: 319-005-CL) supplemented with 10% fetal bovine serum (FBS) (Wisent cat: 080-150). OVCAR8 (RRID:CVCL_1629 ) cells were cultured in high-glucose Roswell Park Memorial Institute's (RPMI) medium containing HEPES buffer, penicillin, and streptomycin (Wisent cat: 350-700-CL) supplemented with 10% FBS (Wisent cat: 080-150). SUM149PT (RRID:CVCL_3422 ), cells were cultured in Ham's F-12 (F12) medium mixture (Gibco cat: 31765035) supplemented with 5% FBS, 10 mM HEPES (Gibco cat: 15630080) 1 μg/ml hydrocortisone (Sigma cat: H0888) and 5 μg/ml insulin (Sigma cat: I-1882). MCF10A (RRID:CVCL_0598 ), cells were grown in a DMEM/F12 medium mixture (Gibco cat: 11320033) supplemented with 5% horse serum (Invitrogen: 16050-122), 20 ng/ml epidermal growth factor (EGF), 100 ng/ml Cholera toxin, 0.5 mg/ml hydrocortisone (Sigma cat: H0888) and 10 μg/ml insulin (Sigma cat: I-1882). Cell lines were passaged using 0.05% Trypsin-EDTA solution (Wisent cat: 325-542-EL) and grown in a humidified atmosphere at 37°C and 5% CO2.
6
Culturing HEK 293T, Vero, and SH-SY5Y Cell Lines
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HEK 293T cells [28 (link),29 (link)] were maintained in Dulbecco’s modified Eagle’s medium (DMEM) with high glucose (Hyclone, Logan, UT, USA) supplemented with 10% fetal bovine serum (FBS) (Hyclone, Logan, UT, USA), 50 U/mL Penicillin and 50 μg/mL Streptomycin (Thermo Fisher Scientific, Burlington, ON, Canada). African green monkey epithelial Vero cells [30 (link)] were maintained in DMEM with high glucose, supplemented with 5% FBS, 1% non-essential amino acids, 1% L-glutamine, 50 U/mL Penicillin and 50 μg/mL Streptomycin (Wisent, St Bruno, QC, Canada). Neuroblastoma SH-SY5Y cells [31 (link)] were obtained from Dr. Marc Fabian (LDI and McGill University, Montréal, QC, Canada) and maintained in DMEM/Ham’s F12 50/50 mix (Wisent, St Bruno, QC, Canada), supplemented with 10% FBS, 50 U/mL Penicillin and 50 μg/mL Streptomycin.
7
Differentiation of Mouse Bone Marrow-Derived Immune Cells
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For the differentiation of bone marrow derived macrophages (BMDMs), 4 x106 mouse bone marrow cells were plated into 5 mLs of DMEM (Wisent) with 20% FCS, 2mM L-Glutamine, 1mM sodium pyruvate, 100μg/ml streptomycin, 100U/ml penicillin, 1μg/ml amphotericin B (all from Wisent), and 25% L929 cell conditioned media (prepared in house according to standard protocols, contains M-CSF). For the differentiation of bone marrow derived dendritic cells (BMDCs), 4 x106 mouse bone marrow cells were plated into 5 mLs of RPMI-1640 (Wisent) with 10% FCS, 2mM L-Glutamine, 1mM pyruvate, 100μg/ml streptomycin, 100U/ml penicillin, 1μg/ml amphotericin B (all from Wisent), 50 μM β-mercaptoethanol (Calbiochem), and 25 ng/mL GM-CSF (Peprotech). The cells were maintained at 37°C and 5% CO2 in a humidified incubator for 1 week. To detach, the cells were washed in PBS (Wisent) and treated with Versene (0.02% w/w EDTA in PBS, Wisent). Successful derivation of macrophages and dendritic cells was confirmed by the changing morphology of the cells and by flow cytometry analysis for the expression of CD11b, F4/80 and CD11c lineage markers.
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Immortalized murine macrophage culture
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Immortalized bone marrow-derived macrophages from B10A.Bcgr mice (B10R macrophages) were grown and maintained at 37°C in 5% CO2 in Dulbecco’s Modified Eagle’s Medium (Wisent Inc., St-Bruno, QC) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Invitrogen, Burlington, ON, Canada), 2mM L-glutamine, 100U/ml penicillin, and 100 μl/ml streptomycin (Wisent, St-Bruno, QC, Canada). BHK-21 cells (ATCC) were grown and maintained at 37°C with 5% CO2 in Eagle’s Minimum Essential Medium (Wisent) supplemented with 5% heat-inactivated FBS, 2mM L-glutamine, 100 U/ml penicillin, and 100 μl/ml streptomycin.
Primary antibodies purchased for immunoblotting were phospho-p44/42 (ERK 1/2) (#9101S); ERK 1/2 (#4695S); phospho-p38 (#9216S); p38 (#9212S); phospho-IRF3 (4D4G); IRF3 (D6I4C), phospho-TBK1/NAK (D52C2), and TBK (D1B4) (Cell Signaling Technology, Danvers, MA, USA).
Primary antibodies purchased for immunoblotting were phospho-p44/42 (ERK 1/2) (#9101S); ERK 1/2 (#4695S); phospho-p38 (#9216S); p38 (#9212S); phospho-IRF3 (4D4G); IRF3 (D6I4C), phospho-TBK1/NAK (D52C2), and TBK (D1B4) (Cell Signaling Technology, Danvers, MA, USA).
9
Murine Hematopoietic Progenitor Cell Culture
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Murine B cell progenitor cell line Ba/F3 (DSMZ, ACC 300) was maintained at 0.5–2 × 106 cells/mL in RPMI-1640 (Wisent) with 10% fetal calf serum (FCS, Wisent), 2 mM L-Glutamine, 100 μg/mL streptomycin, 100 U/mL penicillin (Wisent), and 5% WEHI conditioned media as the source of interleukin-3. Multipotent hematopoietic progenitor cells HPC7, derived from murine ES-cells via constitutive expression of LIM-homeobox gene LH2, were provided by Professor Leif Carlsson (Umea Center for Molecular Medicine, Umea, Sweden) [27 (link)] and previously extensively characterized [72 (link)]. HPC7 cells were cultured at 0.5–2 × 106 cells/mL in IMDM (Life Technologies), with 10% FCS (Gibco, Life Technologies), 100 μg/mL streptomycin, 100 U/mL penicillin (Wisent), 7.48 × 10−5 M MTG (M6145, Sigma-Aldrich), and 100 ng/mL murine stem cell factor (Shenandoah Biotechnology, Cedarlane).
10
Mammalian Cell Culture Protocols
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