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Antibiotic antifungal

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The Antibiotic/antifungal is a laboratory product designed to test the effectiveness of antibiotics and antifungal agents against microbial samples. It provides a standardized method to determine the minimum inhibitory concentration (MIC) of these compounds.

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Lab products found in correlation

Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions) Dulbecco s minimal essential medium dmem, by Corning (2 mentions) Hek293 lentix cells, by Takara Bio (2 mentions) Non essential amino acids (neaa), by Thermo Fisher Scientific (2 mentions) Minimum essential medium (mem), by Corning (2 mentions) 0.22 μm filter, by Celltreat (2 mentions) Trypsin, by Thermo Fisher Scientific (1 mentions) Amplex red cholesterol assay kit, by Thermo Fisher Scientific (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Fluoromount g, by Southern Biotech (1 mentions) Sodium pyruvate, by Thermo Fisher Scientific (1 mentions) Anti mouse cd3, by BD (1 mentions) Soluble anti mouse cd28, by BD (1 mentions) Mem non essential amino acid, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Hepes buffer, by Thermo Fisher Scientific (1 mentions) Rpmi 1640, by Cytiva (1 mentions) Rnalater solution, by Thermo Fisher Scientific (1 mentions)

7 protocols using antibiotic antifungal

1

Propagation of SVG-A and HEK293-LentiX Cells

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SVG-A is a transformed cell line derived from primary fetal human glial cells and was described previously (Major et al., 1985 (link)). SVG-A cells were grown in minimal essential medium (MEM) (Corning, Inc, New York, NY) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Flowery Branch, GA) and 1% antifungal/antibiotic (Gibco Life Technologies, Gaithersburg, MD). HEK293-LentiX cells are a subclone of the HEK293 line optimal for lentiviral packaging systems and were obtained from (Takara). LentiX cells were grown in Dulbecco’s minimal essential medium (DMEM) (Corning) supplemented with 10% FBS, 1% nonessential amino acids (Gibco Life Technologies), and 1% antifungal/antibiotic. EV-depleted media was used as needed for EV-related experiments while complete media was used for general passage of cell lines. EV-depleted media was produced as described (Théry et al., 2002 ). Briefly, 2X media was prepared and spun at 100,000 ×g in a Type 45 Ti rotor (k-factor = 133), for 18 hours at 4°C. Media was then diluted before use to 1X and filtered through 0.22 μm filter (CELLTREAT, Pepperell, MA). Cells were grown in a humidified chamber at 37°C and 5% CO2.
Generation of the JC polyomavirus strain Mad-1/SVEΔ and production and purification of viral stocks was previously described (Liu & Atwood, 2000 , Liu et al. ).
Morris-Love J., O’Hara B.A., Gee G.V., Dugan A.S., O’Rourke R.S., Armstead B.E., Assetta B., Haley S.A, & Atwood W.J. (2022). Biogenesis of JC polyomavirus associated extracellular vesicles. Journal of extracellular biology, 1(5), e43.
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2

Propagation of SVG-A and HEK293-LentiX Cells

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SVG-A is a transformed cell line derived from primary fetal human glial cells and was described previously (Major et al., 1985 (link)). SVG-A cells were grown in minimal essential medium (MEM) (Corning, Inc, New York, NY) supplemented with 10% fetal bovine serum (FBS) (Atlanta Biologicals, Flowery Branch, GA) and 1% antifungal/antibiotic (Gibco Life Technologies, Gaithersburg, MD). HEK293-LentiX cells are a subclone of the HEK293 line optimal for lentiviral packaging systems and were obtained from (Takara). LentiX cells were grown in Dulbecco’s minimal essential medium (DMEM) (Corning) supplemented with 10% FBS, 1% nonessential amino acids (Gibco Life Technologies), and 1% antifungal/antibiotic. EV-depleted media was used as needed for EV-related experiments while complete media was used for general passage of cell lines. EV-depleted media was produced as described (Théry et al., 2002 ). Briefly, 2X media was prepared and spun at 100,000 ×g in a Type 45 Ti rotor (k-factor = 133), for 18 hours at 4°C. Media was then diluted before use to 1X and filtered through 0.22 μm filter (CELLTREAT, Pepperell, MA). Cells were grown in a humidified chamber at 37°C and 5% CO2.
Generation of the JC polyomavirus strain Mad-1/SVEΔ and production and purification of viral stocks was previously described (Liu & Atwood, 2000 , Liu et al. ).
Morris-Love J., O’Hara B.A., Gee G.V., Dugan A.S., O’Rourke R.S., Armstead B.E., Assetta B., Haley S.A, & Atwood W.J. (2022). Biogenesis of JC polyomavirus associated extracellular vesicles. Journal of extracellular biology, 1(5), e43.
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3

Maqui Berry Polyphenols Modulate Lipid Metabolism

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Culture medium Roswell Park Memorial Institute (RPMI) 1640, Dulbecco’s modified Eagle culture medium (DMEM), fetal bovine serum (FBS), antibiotic-antifungal, phosphate-buffered saline (PBS), trypsin, and ethylenediaminetetraacetic acid (EDTA) were purchased from Gibco™ (Waltham, MA, USA). Nile Red (C20H18N2O2), Oil Red O (C26H24N4O), dimethylsulfoxide (DMSO), Philippine, and paraformaldehyde were purchased from Sigma-Aldrich (St-Louis, MO, USA). Trizol reagent was obtained from Ambion® (Waltham, MA, USA). Fluoromount-G was purchased from Southern Biotech (Birmingham, AL, USA & Canada), and Amplex Red Cholesterol Assay Kit was obtained from Invitrogen™ (Eugene, OR, USA). Delphinidin-3-sambubioside-5-glucoside (DS) and delphinidin-3,5-diglucoside (DG) were isolated from Maqui berries. Olanzapine Zyprexa® injectable solution 10 mg/mL from Laboratorios Eli lilly was used.
del Campo A., Salamanca C., Fajardo A., Díaz-Castro F., Bustos C., Calfío C., Troncoso R., Pastene-Navarrete E.R., Acuna-Castillo C., Milla L.A., Villarroel C.A., Cubillos F.A., Aranda M, & Rojo L.E. (2021). Anthocyanins from Aristotelia chilensis Prevent Olanzapine-Induced Hepatic-Lipid Accumulation but Not Insulin Resistance in Skeletal Muscle Cells. Molecules, 26(20), 6149.
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4

Naive CD4+ T Cell Isolation and Culture

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Treat C57BL/6 mouse spleen tissue, crush on a cell filter to obtain a single cell suspension, and lyse erythrocytes. Naïve CD4+ T cells were sorted according to the MagniSort™Mouse CD4 Naïve T cell Enrichment Kit supplier time instructions (Invitrogen, Carlsbad, CA, USA).
For all T cell cultures, RPMI 1640 (Cytiva, Marlborough, MA, USA) medium was supplemented with 10% FBS (GIBCO, Grand Island, NY, USA), 10 mM HEPES buffer (GIBCO, Grand Island, NY, USA), 55 mM 2-ME (GIBCO, Grand Island, NY, USA), antibiotic–antifungal (GIBCO, Grand Island, NY, USA) and 2 mM L -glutamine (GIBCO, Grand Island, NY, USA), 1 mM sodium pyruvate (GIBCO, Grand Island, NY, USA) and MEM non-essential amino acids (GIBCO, Grand Island, NY, USA). About 100,000 cells were added to 96-well plates coated with anti-mouse CD3 (2.5 mg/mL, BD Pharmingen, San Diego, CA, USA) and soluble anti-mouse CD28 (2 mg/mL, BD Pharmingen, San Diego, CA, USA). T cells were cultured with CCL2 or CCL8 (MedChem Express, Monmouth Junction, NJ, USA).
Jin R., Zhou M., Lin F., Xu W, & Xu A. (2023). Pathogenic Th2 Cytokine Profile Skewing by IFN-γ-Responding Vitiligo Fibroblasts via CCL2/CCL8. Cells, 12(2), 217.
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5

Placental Umbilical Tissue Biopsy and Newborn Blood Sampling

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Umbilical tissue biopsies were obtained once the placenta was delivered and used for immunohistochemical, genetic and molecular studies. The biopsy fragments were introduced into 2 different sterile tubes: One containing Minimum Essential Medium (MEM) with 1% antibiotic/antifungal (both from Thermo Fisher Scientific, Waltham, MA, USA) and another containing RNAlater® solution (Ambion, Austin, TX, USA). In the laboratory, the samples were processed in a Class II Telstar AV 30/70 Müller 220 V 50 MHz laminar air flow cabinet (Telstar SA Group, Terrassa, Spain) to maintain a sterile environment.
The preserved samples were kept in 1 mL of RNAlater® at −80 °C until processing for gene expression analysis. The samples preserved in MEM were reserved for histological and immunohistochemical studies.
Blood samples were obtained from the newborns by umbilical vein puncture after delivery. Plasma was obtained from these blood samples for the MDA study. The total volume of sampled blood was transferred from heparinized tubes to sterile centrifuge tubes, which were centrifuged at 1500× g for 15 min. Next, the plasma was collected and transferred to 1.5 mL Eppendorf tubes, where it was kept at −80 °C until it was used for the study.
Ortega M.A., Sánchez-Trujillo L., Bravo C., Fraile-Martinez O., García-Montero C., Saez M.A., Alvarez-Mon M.A., Sainz F., Alvarez-Mon M., Bujan J., De Leon-Luis J.A, & García-Honduvilla N. (2021). Newborns of Mothers with Venous Disease during Pregnancy Show Increased Levels of Lipid Peroxidation and Markers of Oxidative Stress and Hypoxia in the Umbilical Cord. Antioxidants, 10(6), 980.
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6

Culturing Liver Cancer Cell Lines

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The liver cancer cell lines HepG2, Hep3B and SNU-387 (HB-8065) were used (ATCC, Manassas, USA). HepG2 and Hep3B cells were cultured in minimum essential medium (Thermo Fisher, Madrid, Spain) supplemented with 10% fetal bovine serum (FBS; Sigma‒Aldrich, Madrid, Spain), 0.2% antibiotic-antifungal (Gentamicin/amphotericin-B, Thermo Fisher) and 0.5% sodium pyruvate8 (link). SNU-387 cells were cultured in Roswell Park Memorial Institute 1640 (RPMI-1640; Thermo Fisher) medium supplemented with 10% FBS, 0.2% antibiotic-antifungal and 0.5% glutamine (Thermo Fisher)8 (link). Cells were maintained at 37 °C and 5% CO2 under sterile conditions, periodically validated by short tandem repeat analysis (GenePrint 10 System, Promega, Barcelona, Spain) and tested for mycoplasma contamination8 (link),17 (link),18 (link). Positive and/or negative controls [IGF-1 (10-6 M) and paclitaxel (10-7 M), respectively] were used.
López-Cánovas J.L., Hermán-Sánchez N., del Rio-Moreno M., Fuentes-Fayos A.C., Lara-López A., Sánchez-Frias M.E., Amado V., Ciria R., Briceño J., de la Mata M., Castaño J.P., Rodriguez-Perálvarez M., Luque R.M, & Gahete M.D. (2023). PRPF8 increases the aggressiveness of hepatocellular carcinoma by regulating FAK/AKT pathway via fibronectin 1 splicing. Experimental & Molecular Medicine, 55(1), 132-142.
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7

Decellularization and Digestion of Tissue for pECM Production

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The decellularization and digestion protocols were developed based on previous studies28 (link),30 (link). The minced tissues were treated with 1% (v/v) antibiotic/antifungal (Thermo Fisher Scientific) and placed on a shaker overnight at room temperature. The tissue was then treated in 0.1% Sodium Dodecyl Sulfate (SDS) in distilled water for 48 hours to decellularize the tissue without any buffer change. Following SDS treatment, the tissue is washed in distilled water for another 48 hours and lyophilized. The lyophilized, decellularized tissue was crushed using a mortar and pestle set and digested in pepsin (1 g of pepsin / 10 g of dry tissue) in 0.5M acetic acid in 4°C for 48 hours. The pH of the tissue digest was adjusted to 7.4 using an aqueous base solution (e.g. NaOH) and then centrifuged at 3000 g for 5 mins. The supernatant was filtered using a cell strainer (100 μm, Thermo Fisher Scientific) and stored at −80°C until further use. The resulting supernatant was defined as pECM for the current work. The DNA content of pECM was tested using a Quant-iT PicoGreen dsDNA Assay kit (Life Technologies) following the manufacturer’s protocol. Collagen content of pECM was tested using a hydroxyproline assay kit (Sigma-Aldrich).
Kuo C.Y., Guo T., Cabrera-Luque J., Arumugasaamy N., Bracaglia L., Garcia-Vivas A., Santoro M., Baker H., Fisher J, & Kim P. (2018). Placental Basement Membrane Proteins are Required for Effective Cytotrophoblast Invasion in a 3D Bioprinted Placenta Model. Journal of biomedical materials research. Part A, 106(6), 1476-1487.
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