Human colon cancer cell lines: DLD-1 (ATCC: CCL-221TM), LoVo (ATCC: CCL-229TM), HCT8 (ATCC: CCL-244TM), SW480 (ATCC: CCL-228TM) were used. mRNA expression analyses were performed on DLD-1, HCT8, LoVo and SW480 cell lines. HCT8 cells were used in RNAi experiments, proliferation rate and wound healing assay. DLD-1 were employed in all the experiments mentioned in the manuscript. Human colon cancer cell lines and kidney HEK-293T cells were grown in RPMI-1640 (Gibco, Life Technologies Corporation, Carlsbad, CA, USA) and Dulbecco's Modified Eagle Medium (DMEM, GE healthcare, Milan, IT), respectively. RPMI-1640 was supplemented with 15% fetal bovine serum (FBS) (Euroclone, Milan, IT), Glutamine (Euroclone, Milan, IT), non-essential Amino Acids (Gibco, Life Technologies Corporation, Carlsbad, CA, USA), Penicillin-Streptomycin (Gibco, Life Technologies Corporation, Carlsbad, CA, USA). DMEM medium was completed with 10% FBS, Glutamine, Penicillin-Streptomicin, (Euroclone, Milan, IT). Human colon cancer cell lines were kindly gifted by G. Casey (USC Norris Comprehensive Cancer Center, Los Angeles, USA).
Penicillin streptomicin
Manufactured by Euroclone
Sourced in Italy
Penicillin/streptomycin is a commonly used antibiotic solution for cell culture applications. It provides protection against bacterial contamination in cell culture systems.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Euroclone (5 mentions)
L glutamine, by Euroclone (5 mentions)
Rpmi 1640 medium, by Euroclone (3 mentions)
Dulbecco modified eagle medium (dmem), by Euroclone (2 mentions)
Rpmi 1640, by Euroclone (2 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (1 mentions)
Glutamine, by Euroclone (1 mentions)
Dulbecco s modified eagle medium dmem, by GE Healthcare (1 mentions)
Rpmi 1640 medium without l glutamine with phenol red, by Euroclone (1 mentions)
Dmem high glucose, by Euroclone (1 mentions)
Ebm 2 basal medium, by Lonza (1 mentions)
Dmem low glucose, by Euroclone (1 mentions)
Singlequots kit, by Lonza (1 mentions)
Huvecs, by Merck Group (1 mentions)
Mg 63, by Merck Group (1 mentions)
Ecacc, by Merck Group (1 mentions)
Saos 2, by Merck Group (1 mentions)
Huvecs, by Lonza (1 mentions)
L glutamine, by Lonza (1 mentions)
10 protocols using penicillin streptomicin
1
Comparative Analysis of Colon Cancer Cell Lines
2
Differentiation of THP-1 Macrophages
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THP-1 and THP-1 X-Blue™ cells were maintained in RPMI 1640 Medium without L-glutamine with phenol red (Euroclone, Pero, Italy), supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Euroclone, Pero, Italy), 2 mM L-glutamine (Euroclone, Pero, Italy), and 100 U/mL penicillin/streptomicin (Euroclone, Pero, Italy). Before treatments, cells were seeded into a 96-well plate and differentiated into macrophages by 72 h incubation with 100 ng/mL phorbol 12-myristate 13-acetate (PMA, Enzo Life Sciences, New York, NY, USA) followed by 24 h incubation in RPMI medium.
3
Generating PLX4032-Resistant Melanoma Cell Line
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MEOV NT cell line, directly obtained from the biopsy of an untreated patient with metastatic melanoma (MM), was kindly provided by Prof. Gabriella Pietra (University of Genoa, Genoa, Italy). The PLX4032-resistant cell line (MEOV PLX-R) was selected by treating MEOV NT cells for 6 months with increasing concentrations of PLX4032 [25 (link)]. Briefly, the drug-resistant population was obtained by seeding the MEOV NT cells twice a week at a density of 1.5 × 106 and treated over six months with PLX4032 (250 nM–1.5 µM). The authenticity of the selected cells was checked by Short Tandem Repeat (STR) profile analysis performed by the Immunohematology and Transfusion operative unit, IRCCS Ospedale Policlinico San Martino, Genoa. Both cell lines were maintained in RPMI 1640 medium (Euroclone Spa, Pavia, Italy) supplemented with 10% Fetal Bovine Serum (FBS, Euroclone Spa, Pavia, Italy), 1% L-Glutamine (Euroclone Spa, Pavia, Italy), and 1% Penicillin/Streptomicin (Euroclone Spa, Pavia, Italy) and grown in standard conditions (37 °C humidified incubator with 5% CO2).
4
Culturing MG63 Osteosarcoma Cells
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The human osteosarcoma cell line MG63, a well-established cell model, was obtained from the American Type Culture Collection (ATCC, Rockville, MD, USA). MG63 osteoblast-like cells were cultured and maintained in a monolayer in T25 cm2 culture flasks in High Glucose Dulbecco’s Modified Eagle’s Medium (DMEM High Glucose, Euroclone, Milan, Italy) supplemented with 10% fetal calf serum (FCS) and 1X Penicillin-Streptomicin (Euroclone), at 37 °C in a humified 5% CO2 incubator.
5
Experimental Compound Dissolution and Cell Culture
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The experimental compounds herein tested were dissolved in DMSO at a stock concentration of 15 mM, aliquoted and frozen at –20°C. The aliquot in use was kept at room temperature (rt). Control cells always contained DMSO at a concentration corresponding to that of the highest concentration of the compounds.
Cells were cultured in complete medium consisting of RPMI 1640 or DMEM (the latter for fusion assays) (Euroclone) supplemented with antibiotics penicillin/streptomicin and glutamin plus 10% heat-inactivated fetal calf serum (FCS) (Euroclone).
Maraviroc and AMD3100 were obtained from the Centre for AIDS Reagents, National Institute for Biological Standards & Control (NIBSC), UK (referred to as NIBSC, UK).
Cells were cultured in complete medium consisting of RPMI 1640 or DMEM (the latter for fusion assays) (Euroclone) supplemented with antibiotics penicillin/streptomicin and glutamin plus 10% heat-inactivated fetal calf serum (FCS) (Euroclone).
Maraviroc and AMD3100 were obtained from the Centre for AIDS Reagents, National Institute for Biological Standards & Control (NIBSC), UK (referred to as NIBSC, UK).
6
Culturing Human Cell Lines for Research
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Human OS cell lines (SaOS‐2, MG‐63 and HOS) and human umbilical vein endothelial cells (HUVECs) were purchased from Sigma Aldrich (ECACC). Tumour cells were maintained in Dulbecco's modified Eagle's medium (DMEM) high glucose with 2 mm l ‐glutamine (Euroclone, Milan, Italy) supplemented with fetal bovine serum (FBS; 10% v/v, Euroclone) and penicillin/streptomicin (P/S, 1% v/v, Euroclone), in cell culture flasks until 70–80% cell confluence. HUVECs were cultured in complete endothelial cell growth medium (EBM‐2 Basal Medium, Lonza, Basel, Switzerland) plus the SingleQuots Kit, supplemented with 2 mm l ‐glutamine, P/S (1% v/v) and FBS (10% v/v). Human BM‐MSCs used in this study were isolated from healthy donors and characterized by Dr Bambi's Unit (AOU Meyer Hospital, Florence, Italy) as previously reported (Barcellos‐de‐Souza et al., 2016 ). BM‐MSCs were cultured in DMEM low glucose (Euroclone) containing FBS (10% v/v), 2 mm l ‐glutamine and P/S (1% v/v) and used between passages 2 and 7.
7
Metastatic Melanoma Cell Characterization
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All the experiments were performed on MeOV (BRAFV600E) and MeTA (BRAFV600D) metastatic melanoma cell lines isolated from the biopsies of TT-untreated patients (17 (link)). Melanoma cells were maintained in RPMI 1640 medium (Euroclone Spa, Pavia, Italy) supplemented with 10% Fetal Bovine Serum (FBS, Euroclone Spa, Pavia, Italy), 1% L-Glutamine (Euroclone Spa, Pavia, Italy) and 1% Penicillin/Streptomicin (Euroclone Spa, Pavia, Italy) and grown under standard conditions (37°C humidified incubator with 5% CO2).
8
Establishing Drug-Resistant NB Cell Lines
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HTLA-230 human stage-IV NB cells were kindly provided by Dr. L. Raffaghello (G. Gaslini Institute, Genoa, Italy). The HTLA-ER cells were selected by treating HTLA-230 parental cells for six months with increasing concentration of etoposide, as previously reported [34 (link)]. Both cell populations were maintained in RPMI 1640 medium (Euroclone Spa, Pavia, Italy) supplemented with 10% Fetal Bovine Serum (FBS, Euroclone Spa, Pavia, Italy), 1% L-Glutamine (Euroclone Spa, Pavia, Italy) and 1% Penicil-lin/Streptomicin (Euroclone Spa, Pavia, Italy) and grown in standard conditions (37 °C humidified incubator with 5% CO2).
9
Optimized Breast Cancer Cell Culture for EV Isolation
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Cell cultures and experimental conditions for small extracellular vesicle isolation MCF7 and MDA-MB-231 breast cancer cell lines were cultivated in DMEM (Dulbecco's Modified Eagle's Medium High Glucose with Sodium Pyruvate with L-Glutamine, EuroClone, ECM0728L) supplemented with 10 % FBS (Fetal Bovine Serum South America origin EU, EuroClone, ECS0180L) and 1 % Penicillin/Streptomicin (100×, EuroClone, ECB3001D). Cell lines were grown at 37 °C in humidified 5 % CO 2 incubator and split every 2-3 days according to their confluence. The culture and harvesting conditions, such as passage number and seeding confluence, were maintained the same and regular checks for Mycoplasma contamination were performed on cells for vesicle isolation.
10
Primary Mouse Cortical Neuron Culture
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Primary cultures of mouse cortical neurons derived from C57BL/6 mice (Charles River Italia, Calco, Italy). All animal studies were approved by the Animal Research Committees of the University of Brescia and follow the Directive 2010/63/EU of the European Parliament and of the Council of 22 September 2010 on the protection of animals used for scientific purposes. The neurons were obtained from cortices of 15-day embryonic mice as previously described (Lanzillotta et al., 2013) . The cells were seeded at the density 2.5 Â 10 5 cells/well in 24-well plates (Nunc-Thermo Scientific, Waltham, MA, U.S.A.) coated with 10 μg/mL poly-l-lysine (Sigma-Aldrich, Saint Louis, MO, U.S.A.) using Neurobasal medium (Invitrogen Corporation, Carlsbad, CA, USA) supplemented with 2% B27 (Invitrogen Corporation), 0.5 mM l-glutamine (Euroclone, Celbio spa, Milan, Italy) and 50 U/mL penicillin/streptomicin (Euroclone). The neurons were used after 11-12 days in vitro (DIV).
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