Cellular senescence was induced in proliferative UC-MSCs using two models: (1) DDIS: the UC-MSCs were cultured with DMEM supplemented with 10% (v/v) FBS, 1% (v/v) penicillin, 1% (v/v) streptomycin (all from Sigma-Aldrich, Madrid, Spain) and 1-μM Etoposide (MedChemexpress, New Jersey, USA) (Table A1 ) for two days. The cells were then washed with PBS and cultured with DMEM supplemented with 10% (v/v) FBS, 1% (v/v) penicillin and 1% (v/v) streptomycin (all from Sigma-Aldrich, Madrid, Spain) for four days (Figure 2 A). (2) TIS: the UC-MSCs were cultured with DMEM supplemented with 10% (v/v) FBS, 1% (v/v) penicillin, 1% (v/v) streptomycin (all from Sigma-Aldrich, Madrid, Spain) and 1-μM Palbociclib (CDK4/6 inhibitor) (MedChemexpress, New Jersey NJ, USA) (Table A1 ) for three days. After that, the cells were washed with PBS and cultured with DMEM supplemented with 10% (v/v) FBS, 1% (v/v) penicillin and 1% (v/v) streptomycin (all from Sigma-Aldrich, Madrid, Spain) for three days. The cells were then washed with PBS and cultured with DMEM supplemented with 10% (v/v) FBS, 1% (v/v) penicillin and 1% (v/v) streptomycin (all from Sigma-Aldrich, Madrid, Spain) for three days (Figure 2 A).
Streptomycin
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Streptomycin is a laboratory product manufactured by Merck Group. It is an antibiotic used in research applications.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Merck Group (6938 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1971 mentions)
L glutamine, by Merck Group (1161 mentions)
Fetal bovine serum (fbs), by Merck Group (1003 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (737 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (477 mentions)
Amphotericin b, by Merck Group (394 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (380 mentions)
Penicillin, by Thermo Fisher Scientific (359 mentions)
Rpmi 1640 medium, by Merck Group (354 mentions)
Streptomycin, by Thermo Fisher Scientific (335 mentions)
Penicillin g, by Merck Group (308 mentions)
Rpmi 1640, by Merck Group (267 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (261 mentions)
Rpmi 1640, by Thermo Fisher Scientific (243 mentions)
L glutamine, by Thermo Fisher Scientific (240 mentions)
Glutamine, by Merck Group (235 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (196 mentions)
Sodium pyruvate, by Merck Group (185 mentions)
Fetal calf serum (fcs), by Merck Group (183 mentions)
8 260 protocols using streptomycin
1
Senescence Induction in UC-MSCs
2
Cell Line Cultivation Protocols
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NIH:OVCAR-3 cell line was grown in RPMI-1640 culture medium (Sigma-Aldrich, Germany), supplemented with 2 mM L-glutamine, 100 U/mL penicillin/100 µg/mL streptomycin (Sigma-Aldrich), 15% fetal bovine serum (FBS) (Sigma-Aldrich, Germany), and 0.01 mg/mL bovine insulin (Sigma-Aldrich). A2780 and A2780 Cis cell lines were cultivated in RPMI 1640 medium supplemented with 10% Fetal Bovine Serum (Sigma Aldrich), 2 mM L-glutamine, 1% antibiotics (penicillin + streptomycin) (all reagents from Sigma Aldrich) and with 1 µM cisplatinum in the case of A2780 Cis cell line. Ishikawa cell line was cultivated in Eagle’s Minimal Essential Medium (MEM) supplemented with 10% FBS, 2 mM L-glutamin, 1 mM sodium pyruvate, 1% NEA, and 1% antibiotics [25 (link)]. BJ HEP cell line was cultivated in Eagle’s Minimal Essential Medium (MEM), supplemented with 10% Fetal Bovine Serum (Sigma Aldrich), 2 mM L-glutamine, 1% antibiotics (penicillin + streptomycin), and 1% Non-Essential Aminoacids (NEA) Solution (Sigma Aldrich) [26 (link)]. HUVEC cell line was cultivated in RPMI medium, with 10% Fetal Bovine Serum (Sigma Aldrich), 2 mM L-glutamine, and 1% antibiotics (penicillin + streptomycin) (all reagents from Sigma Aldrich). Cell cultures in the 12th passage were used [27 (link)].
3
Prostate Cancer and Mesenchymal Stem Cell Co-culture
4
Cell Culture Protocols for Diverse Cell Lines
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Human embryonic kidney (HEK) 293T cells (GenHunter Co., Nashville, TN) were maintained in Dulbecco's Modified Eagle's Medium (DMEM)(Sigma-Aldrich) containing 1.0 g/L glucose, 2 mM L-glutamine (Sigma-Aldrich), 100 IU/mL penicillin (Sigma-Aldrich), 0.1 µg/mL streptomycin (Sigma-Aldrich) and 10% fetal bovine serum (FBS) (HyClone). Three human neuronal cell lines, neuroepithelioma cells (HTB-10, or SK-N-MC), neuroblastoma cells (HTB-11 or SK-N-SH, and SH-SY5Y), and glioblastoma cells (HTB-14; or U-87) (ATCC, Manassas, VA), were cultured in Minimum Essential Medium (Eagle) (MEM) (Sigma-Aldrich) supplemented with 2 mM L-glutamine, 1.0 mM sodium pyruvate, 100 IU/mL penicillin, 0.1 µg/mL streptomycin and 10% FBS. The human embryonic microglial cell line CHME-5 (provided by Dr. Pierre Talbot, Universite du Quebec) was cultured in Dulbecco's Modified Eagle's Medium (DMEM) (Sigma-Aldrich) containing 4.5 g/L glucose, 2 mM L-glutamine, 100 IU/mL penicillin, 0.1 µg/mL streptomycin and 10% FBS. Human monocytic cell line U937 (ATCC Manassas, VA), was cultured in RPMI1640 (Sigma- Aldrich) supplemented with 2 mM L-glutamine, 1.0 mM sodium pyruvate, 100 IU/mL penicillin, 0.1 µg/mL streptomycin and 10% FBS.
5
Culturing Human Cell Lines and Isolating Neutrophils
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HMEC-1 cells (ATCC® CRL-3243TM, USA) were cultured in complete MCDB 131 medium (Thermofisher Scientific, Horsham, UK) containing 10% foetal bovine serum (FBS, Lonza, Basel; Switzerland), L-glutamine dissolved in sterile water supplemented with 15.7 mL/L sodium bicarbonate solution (7.5% w/v), 10 ng/mL epidermal growth factor (EGF); 1 μg/mL hydrocortisone, 100 μg/mL penicillin and 0.1 mg/mL streptomycin (all Sigma Aldrich, Dorset, UK). HUVECs were cultured in complete endothelium growth medium (ATCC, Virginia, USA) containing Endothelial Cell Growth Kit-BBE (ATCC, USA). Human colorectal adenocarcinoma cells (Caco-2, ATCC® HTB-37TM, USA) were cultured in high-glucose Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma Aldrich, USA) containing 10% FBS, L-glutamine dissolved in sterile water with 15.7 mL/L sodium bicarbonate solution (7.5% w/v), 100 μg/mL penicillin and 0.1 mg/mL streptomycin (all Sigma Aldrich, USA). Human primary neutrophils were isolated using Percoll density gradient and suspended in RPMI-1640 medium with 0.5% fatty-acid-free BSA, 2 mM L-glutamine, 100 U/mL penicillin and 0.1 mg/mL streptomycin (all Sigma Aldrich, USA).
6
Culturing Human Colorectal Cancer Cell Lines
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The human CRC cell lines SW480, SW620, LoVo, HCT116, HT29, RKO, LS174T, and the normal human colon epithelial cell line NCM460 were purchased from the American Type Culture Collection (Manassas, VA, United States) and were subcultured and preserved. SW480 and SW620 cells were cultured in l-15 medium supplemented with 10% fetal bovine serum (FBS, Gibco) and 100μg/mL streptomycin (Sigma-Aldrich), NCM460, HT-29, HCT116, LS174T and RKO cells were cultured in RPMI1640 medium containing 10% FBS (Gibco) and 100μg/ml streptomycin (Sigma-Aldrich), and LoVo cells were cultured in DMEM medium supplemented with 10% FBS (Gibco) and 100μg/mL streptomycin (Sigma-Aldrich). All these cell lines except SW480 and SW620 (without CO2) were maintained at 37°C in a humidified atmosphere containing 5% CO2.
7
Maintenance of Myeloma and Lymphoma Cell Lines
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The SP2/0-Ag14 and SP2/0-NS3/4A myeloma cell lines (H-2d)22 (link) was maintained in Dulbecco's modifed Eagle's medium medium supplemented with 10% fetal bovine serum (FBS) (Sigma Aldrich, St Louis, MO, USA), 2 mM L-glutamine, 10 mM HEPES, 100 U ml−1 penicillin, 100 μg ml−1 streptomycin, 1 mM nonessential amino acids, 50 μM β-mercaptoethanol and 1 mM sodium pyruvate (Sigma Aldrich). SP2/0-Ag14 cells with stable expression of NS3/4A were maintained in 800 μg geneticin (G418) per ml complete Dulbecco's modifed Eagle's medium.
The EL-4 and EL-4-NS3/4A lymphoma cell lines (H-2b)17 (link) were maintained in RPMI 1640 medium supplemented with 10% FBS, 10 mM HEPES, 1 mM sodium pyruvate, 1 mM nonessential amino acids, 50 μM β-mercaptoethanol, 100 U ml−1 penicillin and 100 μg ml−1 streptomycin (Sigma Aldrich). EL-4 cells with stable expression of NS3/4A were maintained in 800 μg geneticin (G418) per ml complete RPMI 1640 medium.
RMA-S cells (H-2Kd, H-2Dd or H-2Ld) were maintained in RPMI 1640 medium supplemented with 5% FBS, 2 mM L-glutamine, 100 U ml−1 penicillin and 100 μg ml−1 streptomycin (Sigma Aldrich). All cells were grown in a humidified 37 °C and 5% CO2 incubator.
The EL-4 and EL-4-NS3/4A lymphoma cell lines (H-2b)17 (link) were maintained in RPMI 1640 medium supplemented with 10% FBS, 10 m
RMA-S cells (H-2Kd, H-2Dd or H-2Ld) were maintained in RPMI 1640 medium supplemented with 5% FBS, 2 m
8
Characterizing miRNA Regulation in Leukemia
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The human promyelocytic cell line NB4 and human embryonic kidney cell line HEK-293T were cultured in RPMI-1640 medium (Gibco, BRL, UK) supplemented with 10% FCS (Gibco), 50 U/ml penicillin, and 50 mg/ml streptomycin (Sigma, St. Louis, MO, USA) at 37°C in 5% CO2. The human promyelocytic cell line HL-60 was cultured in IMDM medium (Gibco) supplemented with 10% FCS, 50 U/ml penicillin, and 50 mg/ml streptomycin (Sigma) at 37°C in 5% CO2. The lentivirus packaging cell line HEK-293TN was maintained in DMEM medium supplemented with 10% FCS, 50 U/ml penicillin, and 50 mg/ml streptomycin (Sigma). HEK-293T cell line was only used for dual-luciferase reporter assay and packaging lentivirus; while NB4 and HL-60 AML cells were used to identify the function of miR-451, YWHAZ, c-Myc and HDAC3.
9
Propagation of HSV-1 and Bovine Coronavirus in Cell Cultures
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HSV-1 McKrae [27 (link)] and HSV-1-GFP [28 (link)] were cultured in Vero cells in Eagle’s Minimum Essential Medium (EMEM) supplemented with 5% fetal bovine serum (FBS) (GeminiBio, Sacramento, CA, USA), penicillin (100 U/mL), and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc., St. Louis, MO, USA). Vero cells were cultured in EMEM supplemented with 10% fetal bovine serum (FBS) (GeminiBio, USA), penicillin (100 U/mL), and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc., St. Louis, MO, USA) at 37 °C with 5% CO2 in a humidified incubator as we described previously [22 (link),29 (link)].
Bovine coronavirus (BCoV) was the NADL-Nebraska Strain isolated by the National Veterinary Services Laboratories (NVSL) in 1981 [30 (link)]. BCoV was grown on human rectal tumor (Hrt-18G) cells [30 (link),31 (link)], which were maintained in EMEM supplemented with 10% FBS (GeminiBio, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc.) at 37 °C with 5% CO2 in a humidified incubator. The virus was propagated in EMEM supplemented with 2.5 μg/mL trypsin and 2.5 μg/mL pancreatin, 1× insulin–transferrin–selenium (Cat No. 51500-056, GIBCO) [31 (link)].
Bovine coronavirus (BCoV) was the NADL-Nebraska Strain isolated by the National Veterinary Services Laboratories (NVSL) in 1981 [30 (link)]. BCoV was grown on human rectal tumor (Hrt-18G) cells [30 (link),31 (link)], which were maintained in EMEM supplemented with 10% FBS (GeminiBio, USA), penicillin (100 U/mL) and streptomycin (100 μg/mL) (Sigma-Aldrich, Inc.) at 37 °C with 5% CO2 in a humidified incubator. The virus was propagated in EMEM supplemented with 2.5 μg/mL trypsin and 2.5 μg/mL pancreatin, 1× insulin–transferrin–selenium (Cat No. 51500-056, GIBCO) [31 (link)].
10
Cell Culture Conditions for Murine Cancer Cell Lines
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Hepa1‐6 cells were maintained in DMEM (Wako Pure Chemical Industries) supplemented with 10% FBS (Nichirei Bioscience Inc., Tokyo, Japan), 200 U/mL penicillin, 100 μg/mL streptomycin, and 10 mM HEPES (Sigma‐Aldrich Japan) at 37°C in a humidified atmosphere containing 5% CO2. CT26 cells were maintained in RPMI‐1640 medium (Wako Pure Chemical Industries, Osaka, Japan) supplemented with 10% FBS, 200 U/mL penicillin, 100 μg/mL streptomycin, 10 mM HEPES, and 0.05 mmol/L 2‐mercaptoethanol (Sigma‐Aldrich) at 37°C in a humidified atmosphere containing 5% CO2. EG7‐OVA cells were maintained in RPMI‐1640 medium (Wako Pure Industries, Ltd.) supplemented with 10% FBS, 200 U/mL penicillin, 100 μg/mL streptomycin, 10 mM HEPES, 0.05 mmol/L 2‐mercaptoethanol (Sigma‐Aldrich) and 100 μg/mL G418 (Wako Pure Industries, Ltd.) at 37°C in a humidified atmosphere containing 5% CO2. All cell lines were tested to rule out the presence of mycoplasma contamination.
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