Fungizone

Manufactured by Thermo Fisher Scientific

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Fungizone is a parenteral antifungal agent used for the treatment of serious and life-threatening fungal infections. It contains the active ingredient amphotericin B, a polyene antimicrobial compound that binds to ergosterol, a key component of fungal cell membranes, leading to increased permeability and cell death.

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Lab products found in correlation

942 protocols using fungizone

Establishment and Maintenance of Murine Oral and Pancreatic Cancer Cell Lines

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The AT-84 oral cancer cell line is derived from a murine tumor of the oral mucosa that spontaneously arose in a C3H mice, it was originally isolated by Hier et al. [20 (link)] and was kindly provided by Dr Venuti A. and Dr Paolini F. [21 (link)] (Regina Elena National Cancer Institute, Rome, Italy). AT-84 cells were grown in RPMI-1640 medium (Life Technologies, Courtaboeuf, France) supplemented with 10% FCS (PAA), 100 U/mL penicillin (Life Technologies), 100 mg/mL streptomycin (Life Technologies) and 0.1% fungizone (Life Technologies). Cells were grown at 37 °C in a humidified atmosphere with 5% CO₂.
The Hight TF-Panc02 cell line was derived from a pancreatic ductal adenocarcinoma induced in a C57BL/6 mouse¹. This cell line is a subclone of Panc02 cells transfected with pcDNA 6.2-GW/EmGFP-miR mock described previously by Mezouar et al. [22 (link)]. Hight TF-Panc02 were grown in RPMI-1640 medium (Life Technologies) supplemented with 10% FCS (PAA), 100 U/mL penicillin (Life Technologies), 100 mg/mL streptomycin (Life Technologies) and 0.1% fungizone (Life Technologies). The cells were grown at 37 °C in a humidified atmosphere with 5% CO₂.

Haen P., Crescence L., Mege D., Altié A., Dubois C, & Panicot-Dubois L. (2021). Oral Squamous Cell Carcinoma Is Associated with a Low Thrombosis Risk Due to Storage Pool Deficiency in Platelets. Biomedicines, 9(3), 228.

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Alveolar Organoid Formation from Murine Mammary Cells

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Mammary epithelial cells were isolated from 12- to 14-week-old nonparous mice and embedded (200 organoids per well) in Matrigel (BD Biosciences, San Jose, CA) in a 96-well plate, and allowed to solidify for 30 min. Organoid growth medium (150 μl per well, Dulbecco’s modified Eagle’s medium (DMEM)/F12, 5% fetal bovine serum, insulin-transferrin-sodium selenite (ITS) (Sigma Aldrich), gentamycin (50 µg/ml), penicillin/strepomycin, fungizone (Life Technologies, Carlsbad, CA, USA) was overlaid and incubated for 24 h. Alveologenesis medium (DMEM/F12, ITS, gentamycin (50 µg/ml), pen/strep, fungizone) including growth factors (9 nM FGF10 (Life Technologies), 9 nM bFGF, 9 nM TGFa, and 9 nM recombinant mouse prolactin (Sigma Aldrich)) was added, and replaced every third day. Images were captured on the seventh day after seeding.

Pruitt H.C., Metge B.J., Weeks S.E., Chen D., Wei S., Kesterson R.A., Shevde L.A, & Samant R.S. (2018). Conditional knockout of N-Myc and STAT interactor disrupts normal mammary development and enhances metastatic ability of mammary tumors. Oncogene, 37(12), 1610-1623.

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Differentiation of Equine Mesenchymal Cells

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Eight horses were used for differentiation experiments. For tenogenic control cultures, the cells were cultivated in expansion medium with a seeding density of 10,000 cells/cm². The medium was renewed every 2–3 days and the cells were kept in a 5% CO₂ environment at 37°C. To induce differentiation, we followed protocols established by others [22] . Briefly, to induce osteogenic differentiation, the cells were seeded at a density of 3,000 cells/cm² and cultured in DMEM High Glucose with Glutamax (Life technologies), 10% FCS (Life technologies), 0.6% fungizone (Life technologies), 0.1% gentamicin (Life technologies) and freshly added 10 mM glycerol phosphate (Sigma-Aldrich), 0.1 μM dexamethasone (Sigma-Aldrich) and 0.1 mM L-ascorbic acid 2-phosphate (Sigma-Aldrich). To induce adipogenic differentiation cells were seeded at 20,000 cells/cm² and the induction medium consisted of DMEM Glutamax (Life technologies) with 10% FCS (Life technologies), 0.6% fungizone (Life technologies), 0.1% gentamicin (Life technologies) and freshly added 0.1 μM dexamethasone (Sigma-Aldrich), 0.2 mM indomethacin (Sigma-Aldrich), 0.01 mg/ml insulin (Sigma-Aldrich) and 0.5 mM 3 iso-butyl-1-methyl-xanthine (Sigma-Aldrich). The cells were kept for 21 days in a 5% CO₂ environment at 37°C and the differentiation media were refreshed twice a week.

Cadby J.A., Buehler E., Godbout C., van Weeren P.R, & Snedeker J.G. (2014). Differences between the Cell Populations from the Peritenon and the Tendon Core with Regard to Their Potential Implication in Tendon Repair. PLoS ONE, 9(3), e92474.

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Engineered Cartilage Tissue Analogs

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Engineered cartilage tissue analogs (CTAs) were produced as described previously^{24 (link),25}. Briefly, articular cartilage was harvested from juvenile bovine knees (2–6 months old, Research 87, MA), finely minced, and digested overnight (12–16 hours) in DMEM containing collagenase Type II (298U/mL Worthington, NJ). Tissue digests were filtered (70µm pore mesh), washed with PBS containing 200U/mL penicillin, 200µg/mL streptomycin, 5µg/mL Fungizone (PSF, Life Technologies, NY), and centrifuged at 1750 rpm for 15 minutes at 12°C (3X) until collected into a single suspension. Chondrocytes were resuspended at 5×10⁶ cells/mL in complete medium (high glucose DMEM containing 10% FBS, 100U/mL penicillin, 100µg/mL streptomycin, 2.5µg/mL Fungizone, 1% MEM Vitamin Solution (Gibco), 25mM HEPES buffer, 50µg/mL ascorbic acid²⁵). This cell suspension was plated into ultra-low adhesion (polyHEMA coated) 96 well plates (Corning, NY) at 1×10⁶ cells/well, where chondrocytes coalesced within 24 hours to form a CTA²⁵. CTAs were cultured for a minimum of 14–16 weeks in complete medium prior to injury.

Mohanraj B., Meloni G.R., Mauck R.L, & Dodge G.R. (2014). A High Throughput Model of Post-Traumatic Osteoarthritis using Engineered Cartilage Tissue Analogs. Osteoarthritis and cartilage / OARS, Osteoarthritis Research Society, 22(9), 1282-1290.

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Extraction and Conditioning of Degenerative IVD

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All procedures were performed with the approval of the Institutional Review Board of the University of Utah. Disc degeneration was classified using the Pfirrmann method (Table 1). Degenerative IVD tissue was extracted from the lumbar discs of five patients undergoing surgical intervention for axial back pain, and degenerative IVD conditioned media was produced using a previously described method.^{32 (link),40 (link)} Briefly, surgical IVD tissue (containing both nucleus pulposus [NP] and AF tissue) was transferred into a glass petri dish, washed twice in washing medium (DMEM-HG (Life Technologies) supplemented with 1% gentamycin (Gibco), 1% kanamycin (Sigma-Aldrich), and 1% fungizone (Gibco)) and cut into small pieces (~3 mm²). Next, IVD tissue was weighed, and cultured in DMEM-HG supplemented with 50 μg/mL ascorbic acid (Life Technologies), 5 μg/mL gentamicin, and 0.125 μg/mL fungizone at a media to tissue ratio of 3.5 mL/g for 48 hours (37°C and 5% CO₂).^{32 (link),40 (link)} Following culture, degenerative IVD conditioned media from each individual patient were collected and stored separately by patient at −80°C, until time of experiments (up to 3 months).

Stover J.D., Lawrence B, & Bowles R.D. (2020). Degenerative IVD conditioned media and acidic pH sensitize sensory neurons to cyclic tensile strain. Journal of orthopaedic research : official publication of the Orthopaedic Research Society, 39(6), 1192-1203.

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Mammary Organoid Culture and Alveologenesis

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Mammary epithelial cells were isolated from 12-14 week old nonparous mice and embedded (200 organoids/well) in Matrigel (BD Biosciences) in a 96 well plate, and allowed to solidify for 30 min. Organoid growth medium (150 μl/well, DMEM/F12, 5% FBS, ITS (Sigma Aldrich, St Louis, MO), gentamycin (50μg/mL), penicillin/strepomycin, fungizone (Life Technologies, Carlsbad, CA) was overlaid and incubated for 24 hours. Alveologenesis medium (DMEM/F12, ITS, gentamycin (50μg/mL), pen/strep, fungizone) including growth factors (9 nM FGF10(Life Technologies, Carlsbad, CA), 9 nM bFGF, 9 nM TGFa, and 9 nM recombinant mouse Prolactin (Sigma Aldrich, St Louis, MO)) was added, and replaced every third day. Images were captured on the seventh day after seeding.

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C2C12 Myoblast Differentiation Protocol

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C2C12 cells were seeded to 8-well chamber slides coated with poly-ornithine (3 μg/mL) and laminin (10 μg/mL). Proliferating C2C12 myoblasts were maintained in high glucose Dulbecco’s modified eagle medium (DMEM, Life Technologies, Carlsbad, CA) containing 20% fetal bovine serum (Atlas Biologicals, Fort Collins, CO), 0.1% penicillin/streptomycin (Pen/Strep, Life Technologies, Carlsbad, CA), 2 mm glutamine (Life Technologies, Carlsbad, CA), and 0.1% Fungizone (Life Technologies, Carlsbad, CA) at 37°C and 5% CO2. Differentiation of C2C12 myoblasts into myotubes was achieved by seeding cells at a high density (5 × 108 cells/well) in DMEM containing 2% horse serum (Life Technologies, Carlsbad, CA), 0.1% Pen/Strep, 2 mm glutamine, and 0.1% Fungizone.

Brayman V.L., Taetzsch T., Miko M., Dahal S., Risher W.C, & Valdez G. (2021). Roles of the synaptic molecules Hevin and SPARC in mouse neuromuscular junction development and repair. Neuroscience letters, 746, 135663.

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Isolation and culture of porcine VICs

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VICs were isolated from fresh porcine hearts (Hormel) as previously described (24 (link)). Aortic valve leaflets were removed and rinsed in a wash solution containing Earle’s Balanced Salt Solution (Life Technologies) with 1% penicillin-streptomycin (Life Technologies) and 0.5 μg/mL fungizone (Life Technologies). Then, the leaflets were incubated in 250 units/mL collagenase (Worthington) solution for 30 min. at 37°C and vortexed to remove endothelial cells. Cells were then incubated with collagenase solution for 60 min. at 37°C and vortexed. This solution was filtered with a 100 μm cell strainer and centrifuged. The cell pellet was resuspended in growth media consisting of Media 199 with 15% fetal bovine serum (FBS, Life Technologies), 2% penicillin-streptomycin (Life Technologies), and 0.5 μg/mL fungizone (Life Technologies). Cells were cultured at 37°C and 5% CO₂ on tissue culture polystyrene (TCPS) for expansion before experiments. Cells were passaged using trypsin (Life Technologies) digestion. For experiments, media serum level was reduced to 10% FBS to reduce proliferation.

Mabry K.M., Lawrence R.L, & Anseth K.S. (2015). Dynamic stiffening of poly(ethylene glycol)-based hydrogels to direct valvular interstitial cell phenotype in a three-dimensional environment. Biomaterials, 49, 47-56.

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Isolation and Culture of Bovine Chondrocytes

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The fresh stifle joints from young adult cattle (15–24 months old) were obtained from a local abattoir (Bud’s Custom Meats, Riverside, IA). Articular cartilage was harvested from the femur condyle and rinsed in Hank’s Balanced Salt Solution (HBSS, Life Technologies, California, USA) supplemented with 100 U/μl penicillin, 100 μg/ml streptomycin, and 2.5 μg/μl fungizone (Invitrogen Life Technologies, Carlsbad, CA). Full-thickness cartilage samples were minced into fine pieces and then digested overnight with 0.25 mg/ml collagenase type I and pronase E (1:1) (Sigma-Aldrich, St. Louis, MO) dissolved in culture medium in a shaking incubator overnight (0.25 mg/ml each). After isolation, primary chondrocytes were re-plated and cultured in Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s F12 (1:1 mixture) supplemented with 10% fetal bovine serum (Life Technologies, Grand Island, NY), 50 μg/μl L-ascorbate, 100 U/μl penicillin, 100 μg/ml streptomycin, and 2.5 μg/μl fungizone at 37 °C with 5% CO₂.

Yu Y., Moncal K.K., Li J., Peng W., Rivero I., Martin J.A, & Ozbolat I.T. (2016). Three-dimensional bioprinting using self-assembling scalable scaffold-free “tissue strands” as a new bioink. Scientific Reports, 6, 28714.

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Isolation of Cancer-Associated Fibroblasts

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Tumour tissue was minced and transferred into a 10 cm culture dish. 10 ml of RPMI supplemented with 10% FCS, 1% fungizone (Life Technologies), 1% penicillin/streptomycin (Invitrogen) and activated collagenase 1A (1 mg/ml, Sigma) were added and incubated overnight at 37 °C. The digested tissue was then filtered through a cell strainer and centrifuged. The CAF-containing cell pellet was washed in PBS, resuspended in DMEM, 10% FBS, 1% P/S, 1% fungizone, and plated on collagen coated (35 μg/ml, rat tail collagen 1, Gibco) culture dish. Patient samples were obtained through NHS Greater Glasgow and Clyde Biorepository (project #305). All participants gave specific consent to use their tissue samples for research.

Kugeratski F.G., Atkinson S.J., Neilson L.J., Lilla S., Knight J.R., Serneels J., Juin A., Ismail S., Bryant D.M., Markert E.K., Machesky L.M., Mazzone M., Sansom O.J, & Zanivan S. (2019). Hypoxic cancer-associated fibroblasts increase NCBP2-AS2/HIAR to promote endothelial sprouting through enhanced VEGF signaling. Science signaling, 12(567), eaan8247.

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