Fbg from bovine plasma and 2-iminothiolane (2-IT) were purchased from Sigma-Aldrich (Republic of Korea). Doxorubicin hydrochloride was obtained from LC Laboratories (Woburn, MA, USA) and 3-maleimidopropionic acid hydrazonium trifluoroacetic acid (3-MAH) was acquired from Toronto Research Chemicals Inc. (Ontario, Canada). Succinimidyl-[(N-maleimidopropionamido)-tetraethylene glycol] ester (SM(PEG)4) and succinimidyl-[(N-maleimidopropionamid)-dodecaethylene glycol] ester (SM(PEG)12) were purchased from Thermo Scientific (Rockford, IL, USA). Tissue plasminogen activator (t-PA) was acquired from Calbiochem and plasminogen was acquired from EMD Millipore (Darmstadt, Germany). The Celltiter-glo® luminescent cell viability assay kit was obtained from Promega Ltd (Seoul, Republic of Korea). NIH-3T3 and SH-SY5Y cell lines were kindly provided by NanoEntek Inc. (Seoul, Republic of Korea). Gibco® Dulbecco’s Modified Eagle’s Medium (DMEM), Gibco® Roswell Park Memorial Institute (RPMI) 1640, Gibco® penicillin–streptomycin (pen–strep), and Hyclone™ fetal bovine serum (FBS) were purchased from Thermo Fisher Scientific (Seoul, Republic of Korea). All chemicals used in this experiment were of analytical grade. Institutional Review Board approval was not sought for the following research. All principles outlined in the Declaration of Helsinki were followed.
Gibco dulbecco s modified eagle s medium dmem
Manufactured by Thermo Fisher Scientific
Sourced in United States, China
Gibco Dulbecco's modified Eagle's medium (DMEM) is a cell culture medium designed to support the growth and maintenance of various cell lines. It provides a balanced salt solution, amino acids, vitamins, and other nutrients essential for cell proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (10 mentions)
Penicillin and streptomycin, by Thermo Fisher Scientific (2 mentions)
Plasminogen, by Merck Group (1 mentions)
2 iminothiolane 2 it, by Merck Group (1 mentions)
Gibco penicillin streptomycin pen strep, by Thermo Fisher Scientific (1 mentions)
Hyclone fetal bovine serum fbs, by Thermo Fisher Scientific (1 mentions)
Doxorubicin hydrochloride, by LC Laboratories (1 mentions)
Raw264.7 cell, by American Type Culture Collection (1 mentions)
Lo2 cells, by American Type Culture Collection (1 mentions)
B16 cells, by American Type Culture Collection (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Alt activity assay kit, by Merck Group (1 mentions)
Lab tek 2 chamber slide system, by Thermo Fisher Scientific (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions)
Ast activity assay kit, by Merck Group (1 mentions)
Dulbecco s pbs, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Bovine serum albumin (bsa), by Merck Group (1 mentions)
Gibco fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Gibco rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
18 protocols using gibco dulbecco s modified eagle s medium dmem
1
Doxorubicin-Conjugated Fibrinogen Nanoparticles
2
Synthesis and Characterization of Multifunctional Nanoparticles
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HAuCl4·3H2O, sodium citrate, 1-adamantanethiol, Cy5-PEG (2k)-SH, and FITC-PEG (2k)-SH were all procured from Aladdin (China). Zhiyuan Biological Technology Co. Ltd. (China) provided mono(6-mercapto-6-deoxy)-β-CD. Fetal bovine serum (FBS) and Gibco Dulbecco’s modified Eagle’s medium (DMEM) were obtained from Gibco (China). All other chemical reagents were supplied by Aladdin (China).
B16 cells, LO2 cells, and RAW264.7 cells were obtained from American Type Culture Collection (ATCC; USA). All of these cell lines were authenticated by DNA fingerprinting, isozyme detection, viability test, and mycoplasma detection. E. coli (ATCC 33694) was purchased from ATCC. Male C57BL/6 mice (6 weeks) were purchased from Faculty of Health Sciences, University of Macau. All animal procedures were approved by the Animal Ethics Committee, University of Macau and were conducted in accordance with the Animal Management Rules of the Ministry of Health of the P. R. China.
B16 cells, LO2 cells, and RAW264.7 cells were obtained from American Type Culture Collection (ATCC; USA). All of these cell lines were authenticated by DNA fingerprinting, isozyme detection, viability test, and mycoplasma detection. E. coli (ATCC 33694) was purchased from ATCC. Male C57BL/6 mice (6 weeks) were purchased from Faculty of Health Sciences, University of Macau. All animal procedures were approved by the Animal Ethics Committee, University of Macau and were conducted in accordance with the Animal Management Rules of the Ministry of Health of the P. R. China.
3
Multiparameter Antibody and Compound Analysis
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Phycoerythrin (PE)–conjugated anti-human ICAM1 antibody (clone: HCD54), PE-conjugated mouse IgG isotype (PE-IgG), anti-mouse ICAM1 antibody (clone #YN1/1.7.4), and red blood cell (RBC) lysis buffer were purchased from BioLegend (San Diego, CA, USA). PE-conjugated anti-human TROP-2 antibody (clone #77220) and mouse and rat IgG isotype controls were purchased from R&D Systems (Minneapolis, MN, USA). Purified anti-human CD54 Antibody (clone: R6.5), MC-VC-PAB-MMAE, MC-MMAF, Mal-EBE-MaL-DM1, and Mal-EBE-Mal-DM4 were obtained from MabPlex (Yantai, China). Lab-Tek II Chamber Slide System was obtained from Thermo Fisher Scientific (Waltham, MA, USA). Doxorubicin, bovine serum albumin (BSA), AST activity assay kit, ALT activity assay kit, creatinine activity assay kit, and urea activity assay kit were purchased from Sigma-Aldrich (St. Louis, MO, USA). Dulbecco’s PBS, Dulbecco’s PBS, 4′,6-diamidino-2-phenylindole (DAPI), Gibco Dulbecco’s modified Eagle’s medium (DMEM), Gibco DMEM/F12(1:1), and RPMI-1640 medium were purchased from Invitrogen (Carlsbad, CA, USA). The Dojindo Cell Counting Kit CCK-8 was purchased from Dojindo Molecular Technologies (Rockville, MD, USA).
4
Cell Culture Protocols for Cancer Research
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C33A, HeLa, and CaSki cells were purchased from the National Collection of Authenticated Cell Cultures. C33A and HeLa cells were maintained in Gibco® Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Inc), whereas CaSki cells were cultured in Gibco® RPMI 1640 medium (Thermo Fisher Scientific, Inc.). All media were supplemented with 10% Gibco® fetal bovine serum (FBS) (Thermo Fisher Scientific, Inc) and 1% penicillin/streptomycin (Beyotime Institute of Biotechnology). Cells were kept in an incubator in a humidified atmosphere of 5% CO2 at 37°C.
5
Culturing U87 Glioblastoma Neurospheres
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The U87 GBM cell line was purchased from the Chinese Academy of Sciences Cell Bank (Shanghai, China) and cultured in Gibco Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) with 10% fetal bovine serum (FBS). The conditional culture medium, Gibco DMEM/F12 (Thermo Fisher Scientific, Inc.) was supplemented with Invitrogen B27 (1X; Thermo Fisher Scientific, Inc.), 20 ng/ml basic fibroblast growth factor and 20 ng/ml epidermal growth factor (both from PeproTech, Inc., Rocky Hill, NJ, USA). First generation U87-neurospheres were observed in all wells of a 6-well plate 72 h later. All cultures were maintained at 37°C in an atmosphere of 5% CO2.
6
Hepatocellular Carcinoma Tissue Collection
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In total, 36 pairs of HCC and adjacent non-tumor liver tissues were obtained from patients who were pathologically diagnosed with HCC and underwent hepatectomy at Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology (Wuhan, China) between September 2009 and September 2012. Tissue samples were immediately snap frozen in liquid nitrogen until use. The study was approved by the local Research Ethics Committee at Tongji Hospital of Huazhong University of Science and Technology, and written informed consent was obtained from the patients. The characteristics of the included patients are described in Table I .
The cell lines used in this study included the immortalized human fetal liver cell line L02, and eight immortalized human liver cancer cell lines (HepG2, Huh7, PLC/PRF/5, SMMC-7721, SK-HEP-1, MHCC-97H, MHCC-97L and Hep 3B). All were maintained in Gibco Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.), 100 IU/ml penicillin and 100 mg/ml streptomycin, and incubated at 37°C with 5% CO2.
The cell lines used in this study included the immortalized human fetal liver cell line L02, and eight immortalized human liver cancer cell lines (HepG2, Huh7, PLC/PRF/5, SMMC-7721, SK-HEP-1, MHCC-97H, MHCC-97L and Hep 3B). All were maintained in Gibco Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco; Thermo Fisher Scientific, Inc.), 100 IU/ml penicillin and 100 mg/ml streptomycin, and incubated at 37°C with 5% CO2.
7
Culturing diverse cell lines
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Human (Homo sapiens) adenocarcinoma alveolar basal epithelial A549 (American Type Culture Collection [ATCC], Manassas, VA; #CCL-185) cells, epithelial Hela cells (ATCC, #CCL-2), and hepatocarcinoma Huh-7 cells (a kind gift from Hideki Ebihara, Rocky Mountain Laboratories, Hamilton, MT, USA), and grivet (Chlorocebus aethiops) kidney epithelial Vero cells (ATCC, #CCL-81), and Vero E6 cells (ATCC, #CRL-1586) were grown in Gibco Dulbecco’s modified Eagle’s medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% heat-inactivated fetal bovine serum (FBS, Sigma-Aldrich, St. Louis, MO, USA). Syrian golden hamster (Mesocricetus auratus) kidney BHK-21 fibroblasts (ATCC, #CCL-10) were grown in Gibco DMEM supplemented with 10% FBS and 5% tryptose phosphate broth (TPB, Thermo Fisher Scientific). All cells were incubated at 37°C in a humidified 5% CO2 atmosphere.
8
Topical Morin Hydrate for Anti-Inflammatory Effects
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Morin hydrate with 90% of purity was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Lecinolws-50 (lysolecithin; Cosphaderm® E145V)) was purchased from Cosphatec GmbH (Hamburg, Germany). Pluronic F-68 non-ionic surfactant was ordered from Sigma (St Louis, MO, USA). Tween-80 (edge activator) was purchased from Shimakyu’s Pure Chemicals (Osaka, Japan). HaCaT keratinocyte cell line was acquired from the I.Z.S.L.E.R. (Institute Zooprofilattico Sperimentale della Lombardia e dell’Emilia Romagna, Brescia, Italy). Primary antibodies including anti-phospho-ERK 1/2 (Thr202/Tyr204, Thr185/Tyr187) and anti-phospho-p38 (Thr180/Tyr182) were obtained from EMD Millipore Corporation (Temecula, CA, USA). GAPDH antibodies were obtained from Santa Cruz Biotechnology (Dallas, TX, USA) and MMP-1 antibodies were purchased from Proteintech Group, (Rosemont, IL, USA). Fetal Bovine Serum (FBS) and Gibco Dulbecco’s Modified Eagle’s Medium (DMEM) for cell culture were supplied by Thermo Fisher Scientific (Waltham, MA, USA).
9
Cell Line Maintenance Protocols
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All cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA). HCT-15, A549 and MDA-MB-231 cells were maintained in RPMI-1640 medium (Lonza Group, Ltd., Basel, Switzerland). LoVo cells were maintained in F12-K medium (Corning/Cellgro; Corning Inc., Corning, NY, USA). DF1 cells were maintained in Gibco Dulbecco's modified Eagle's medium (DMEM; Thermo Fisher Scientific, Inc., Waltham, MA, USA). All cell culture media were supplemented with 10% fetal bovine serum (FBS; Avantor, Inc., Radnor, PA, USA) and housed in a 5% CO2 atmosphere at 37˚C, except the DF1 cells which were cultured at 39˚C.
10
Isolation and Labeling of Adipose-Derived Stem Cells
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ASCs were obtained from the subcutaneous fat tissue according to a previously described protocol in humans by Zuk et al[31 (link)], based on their adherence to culture, with some minor modifications. Stromal vascular fraction (SVF) was obtained and digested with type I collagenase (0.075%; Gibco®Thermo Fisher Scientific, Waltham, MA, United States). The digested tissue was centrifuged and the pellet was resuspended in 0.16 M NH4Cl for erythrocyte lysis. Then, the cells were separated by filtering the product through a 70-μm nylon mesh, plated in culture dishes, and cultured at 37 °C in a humid atmosphere (90%-95%) with 5% carbon dioxide in the Gibco™ Dulbecco’s Modified Eagle’s Medium (DMEM) (Thermo Fisher Scientific, Waltham, MA, United States) containing 10% phosphate buffered saline (PBS), 2 mmol/L glutamine, and 1% penicillin–streptomycin. Nonadherent cells were removed and subcultured 24 h after seeding; these were the ASCs.
In order to identify ASCs in animal tissue samples, we marked them. In the third subculture at 60% to 70% confluence, ASCs were infected with enhanced green fluorescent protein (eGFP) transducing lentivirus (CNIC, Madrid, Spain). Sorting of selected eGFP-ASCs (99.6% positive) was applied after another 2 to 3 passages.
In order to identify ASCs in animal tissue samples, we marked them. In the third subculture at 60% to 70% confluence, ASCs were infected with enhanced green fluorescent protein (eGFP) transducing lentivirus (CNIC, Madrid, Spain). Sorting of selected eGFP-ASCs (99.6% positive) was applied after another 2 to 3 passages.
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