All MPM cell lines were maintained in a humidified atmosphere containing 5% CO2 in appropriate media supplemented with 10% Fetal Bovine Serum and penicillin streptomycin (500 U/mL). Cell culture reagents were purchased from Lonza (Walkersville, MD, USA). The following MPM cell lines were used in the study: LP9, Met5A, NCI-H2596, MMP, MMB, NCI-H2052, NCI-H28, Ju77, One58, RS-5, DM-3, ACC-MESO-1, ACC-MESO-4, Y-MESO-8D, Y-MESO-9, Y-MESO-12, Y-MESO-14, REN, NCI-H226, and MSTO-211H. ACC-MESO-1, ACC-MESO-4, Y-MESO-9, and Y-MESO-12 were generously provided by Yoshitaka Sekido, (Aichi Cancer Center Research Institute, Japan). NCI-H2052, One-58, and Ju77 cells were provided by Duncan Stewart (University of Leicester, UK). LP-9, MMB, and MMP were a generous gift from Warren Thomas (Royal College of Surgeons in Ireland, Dublin, Ireland). The REN and NCI-H226 cell lines were provided by Dean Fennell (Queen's University, Belfast, Northern Ireland). NCI-H28, and the immortalized non-tumorigenic mesothelial cell line, Met-5A were purchased from the ATCC (LGC Promochem, Teddington, UK). STR profiling of the NCI-H226 was conducted by Source Bioscience (Nottingham, UK) to confirm that the cell line had the correct genotype.
Cell culture reagents
Manufactured by Lonza
Sourced in Switzerland, United States, Belgium
Cell culture reagents are a range of products designed to support the growth and maintenance of cells in a laboratory setting. These reagents provide the necessary components, such as growth media, supplements, and solutions, to create an optimal environment for cell cultivation. The core function of these products is to facilitate the propagation and study of various cell types, enabling researchers to conduct experiments and investigations in a controlled and standardized manner.
Automatically generated - may contain errors
Lab products found in correlation
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions)
Protease inhibitor cocktail, by Merck Group (2 mentions)
Nci h28, by American Type Culture Collection (1 mentions)
Met 5a, by American Type Culture Collection (1 mentions)
Iloprost, by Cayman Chemical (1 mentions)
Daunorubicin hydrochloride, by Merck Group (1 mentions)
Microporous membrane transwell inserts, by Corning (1 mentions)
Calcein am, by Thermo Fisher Scientific (1 mentions)
Doxorubicin hydrochloride, by Merck Group (1 mentions)
Plasmid isolation kit, by Qiagen (1 mentions)
Anti hsp70 antibody, by Thermo Fisher Scientific (1 mentions)
Anti β actin antibody, by Merck Group (1 mentions)
Restriction and dna modifying enzymes, by New England Biolabs (1 mentions)
Htb 55, by American Type Culture Collection (1 mentions)
Infinite m200 pro spectrophotometer, by Tecan (1 mentions)
Milli q ultrapure water, by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole dapi nuclear stain, by Merck Group (1 mentions)
Cytochalasin b, by Merck Group (1 mentions)
Dichloromethane dcm, by Merck Group (1 mentions)
Hplc grade acetonitrile, by Merck Group (1 mentions)
19 protocols using cell culture reagents
1
Maintenance of Mesothelioma Cell Lines
2
Iloprost and Treprostinil Protocol
3
Cell Culture and Pellet Preparation
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Cell culture reagents were obtained from Lonza (Basel, Switzerland), and fetal calf serum from Seromed/Biochrom (Berlin, Germany). All cell lines were grown in IMDM/RPMI (4:1) supplemented with 10% fetal calf serum, glutamine, and 100 U/mL L-penicillin/streptomycin at 37 °C and 5% CO2. For the production of cell pellets, cells were precipitated with pure alcohol, subsequently fixed in 5% formalin solution, and embedded in paraffin.
4
Cytotoxicity and Genotoxicity of Flavonolignans
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The cytotoxicity and genotoxicity experiments were performed on a human lung cancer cell line—A549 (CCL-185; ATCC) cell line, obtained from American Type Culture Collection (ATCC™, Manassas, VA, USA). The cell line was placed in a humidified incubator with a 5% CO2 atmosphere at 37 °C in a DMEM medium, supplemented with 10% (v/v) heat-inactivated Fetal Bovine Serum (FBS), 100 U/mL penicillin, and 100 μg/mL streptomycin. Cell culture reagents were obtained from Lonza (Basel, Switzerland). For cytotoxicity and genotoxicity, cells were seeded at 3 × 106 cells per well and were left overnight before treatment. The following day, the cell samples were incubated with flavonolignans (silybin, silychristin and silydianin), in 3 concentrations (10–50–100 µM) for 24 h.
5
HPLC Analysis of Doxorubicin and Daunorubicin
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Doxorubicin hydrochloride and daunorubicin hydrochloride (internal standard) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Calcein-AM was purchased from Molecular Probes (Eugene, OR, USA). Cell culture reagents were purchased from Lonza group Ltd. (Basel, Switzerland); microporous membrane transwell inserts were purchased from Corning Costar (Acton, MA, USA). Heptanesulfonic acid sodium salt (HPLC-grade) was purchased from TCI Co. Ltd. (Tokyo, Japan). Acetone, ZnSO4 and other reagents, including solvents, were of the highest analytical grade. Deionized water (18.2 MΩ, NANOpure diamond™, Brandstead water purification system, Fistreem International Co. Ltd. (Leicestershire, UK) was used throughout all HPLC analytical steps.
6
Plasmodium Parasite Cell Line Cultivation
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P. falciparum 3D7, D10 parasites expressing ACP-GFP (D10ACP-GFP), and P. berghei ANKA were obtained from the Malaria Research and Reference Reagent Resource Centre (MR4). All the biochemical reagents used in this study were from Sigma and Serva; plasmid isolation kits were from Qiagen or MACHEREY-NAGEL; cell culture reagents were from Lonza and Invitrogen; restriction and DNA modifying enzymes were from New England Biolabs; secondary antibodies and DAPI were from Invitrogen and Thermofisher. Anti-Hsp70 antibody was from Thermo Fisher Scientific (cat No. PA5-11418) and anti-β actin antibody was from Sigma (cat No. A1978-200UL). Human blood was collected from healthy volunteers after written consent under medical supervision at the medical dispensary of the institute, and the protocol (IEC-2/2010) for blood collection for this study has been approved by the Institutional Ethical Committee (IEC) of Centre for Cellular and Molecular Biology.
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P. falciparum 3D7, D10 parasites expressing ACP-GFP (D10ACP-GFP), and P. berghei ANKA were obtained from the Malaria Research and Reference Reagent Resource Centre (MR4). All the biochemical reagents used in this study were from Sigma and Serva; plasmid isolation kits were from Qiagen or MACHEREY-NAGEL; cell culture reagents were from Lonza and Invitrogen; restriction and DNA modifying enzymes were from New England Biolabs; secondary antibodies and DAPI were from Invitrogen and Thermofisher. Anti-Hsp70 antibody was from Thermo Fisher Scientific (cat No. PA5-11418) and anti-β actin antibody was from Sigma (cat No. A1978-200UL). Human blood was collected from healthy volunteers after written consent under medical supervision at the medical dispensary of the institute, and the protocol (IEC-2/2010) for blood collection for this study has been approved by the Institutional Ethical Committee (IEC) of Centre for Cellular and Molecular Biology.
7
Characterization of Calu-3 Lung Cells
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All cell culture reagents were purchased from Lonza Ltd. (Basel, Switzerland) unless stated otherwise. Human lung epithelial cells (Calu-3, ATCC HTB-55™) were selected due to their previous characterisation and study as targets of airborne particulate matter [51 (link),53 (link),69 (link),70 (link),71 (link)]. The immortalised cell line originated from a 25-year-old white male with lung adenocarcinoma. Calu-3 cells were maintained (until passage 20) in Eagle’s minimum essential medium (EMEM) supplemented with 10% (v/v) filter sterilised foetal bovine serum (FBS) (Gibco™, Thermofisher), 1% (v/v) non-essential amino acids, penicillin (50 units/mL), and streptomycin (50 µg/mL) at 37 °C, 5% CO2. Fluorescent and absorbance readings were conducted using a Tecan Infinite M200 pro spectrophotometer.
8
Nedaplatin-Loaded Liposomal Formulation
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Nedaplatin was supplied by Shandong Boyuan Pharmaceutical Co., Ltd. (Shandong, China). DSPC, DSPE, MPEG-2000-DSPE and cholesterol were purchased from Corden Pharma (Plankstdt, Germany). HPLC-grade acetonitrile and dichloromethane (DCM), were purchased from Sigma-Aldrich (Hamburg Germany). Milli-Q ultrapure water was provided using the Millipore system (Watford, UK). For the biological assays all cell culture reagents were purchased from Lonza (Bornem, Belgium), 1-(4,5-Dimethylthiazol-2-yl)-3,5-diphenylformazan (MTT) was purchased from Serva (Heidelberg, Germany), Cytochalasin B and 4′,6-diamidino-2-phenylindole (DAPI) nuclear stain from Sigma-Aldrich (Hamburg, Germany).
9
Culturing Endothelial and Smooth Muscle Cells
10
Cultivation of Various Cell Lines for Virus Research
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Madin-Darby canine kidney (MDCK, American Type Culture Collection) cells were cultivated in EMEM containing 10% fetal bovine serum, 25 mM HEPES, 2 mg/ml bovine serum albumin and antibiotics. DMEM supplemented with 10% fetal bovine serum, 2 mM ultraglutamine and antibiotics was used for A549 (American Type Culture Collection), A549Slam [53] (link) and VeroSlam [54] (link) cells (both supplied by Y. Yanaga, Fukuoka, Japan). A549Slam and VeroSlam stably express the Slam receptor essential for certain measles strains. Non-virally transformed keratinocytes (HaCaT, Cell Lines Services) were grown in DMEM supplemented with 10% fetal bovine serum, 2 mM GlutaMAX I (Invitrogen), 1% modified Eagle’s medium nonessential amino acids and antibiotics. Cell culture reagents were purchased from Lonza unless otherwise indicated.
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