Application Description
Spectrum: Gentamicin Sulfate, EP is a broad-spectrum antibiotic targeting Gram-positive and Gram-negative bacteria. It is effective against several strains of Mycoplasma. It also combats certain β-lactam sensitive VRE or vancomycin resistant Enterococcus; a "superbug."
Microbiology Applications: Gentamicin Sulfate is commonly used as a selective agent to select for cells containing the Gentamicin resistance gene, aacj-AaphD or aacC1. Gentamicin Sulfate is generall used at a concentration of 10 - 50 µg/mL for eukaryotic cell culture and 15 ug/ml for prokaryotic cells.
According to the CDC SOP (DSR-052-04) for Viral Transport Medium (VTM), Gentamicin Sulfate is used at a final concentration of 100 µg/ml.
Media Supplements
Gentamicin can be used as a selective agent in several types of isolation media:
Columbia Blood Agar - Gardnerella vaginalis Selective Supplement
VRE Medium - VRE Selective Supplement
Burkholderia cepacia Agar Base - Burkholderia cepacia Selective Supplement
Plant Biology Applications: Gentamicin sulfate inhbited tracheary element differentiation in cultured explants of pith parenchyma from romaine lettuce (Lactuca sativa L. var. Romana) at 50 to 100 μg/ml. Similar results were obtained with cultured explants of Jerusalem artichoke tuber (Helianthus tuberosus L.). Callus formation was suppressed by Gentamicin Sulfate in both tissue systems. When studying cell division and xylem differentiation in plant tissue culture, concentrations of 10 μg/ml or less is suggested.
Eukaryotic Cell Culture Applications: Gentamicin was found to inhibit Glucose-6-phosphate dehydrogenase (G6PD) enzyme activity in rat erythrocytes (Temel et al, 2018). A bovine macrophage (Bomac) cell line was used but proved to be contaminated with BVDV (bovine viral diarrhea virus). Both infected and cured cells were tested for uptake of Mycoplasma bovis in an in vitro model to dissect the molecular and cellular details of bovine mycoplasmosis using Gentamicin from TOKU-E in a gentamicin protection assay (Burgi et al, 2018). Gentamicin is an effective in vitro bacterial inhibitor that is nontoxic in tissue culture. It was non-toxic to RMK (Rhesus monkey kidney), HeLa, amnion, GMK, and WI-38 cell lines. It can also be used in virus tissue culture as it does not inhibit virus replication. (Rudin et al, 1970). When primary cultures of embryonic rat fibroblasts are exposed to Gentamicin, they develop typical lysosomal phospholipidosis characterized by decreased sphingomyelinase activity, increase in lipid phosphorus, and appearance of 'myeloid bodies' in lysosomes. However, when inhibitors of cysteine proteinases were used (leupeptin, E-64), authors observed a protective effect which could be due to increased sphingomyelinase activity (Montenez et al, 1984).
Cancer Applications: Ovarian melanoma tumor cells was studied in 3D culture and Gentamicin Sulfate was used to prevent contamination when studying ovarian cell lines (OVCAR3, SKOV3, 222, EG, and A2780-PAR ) and normal ovarian surface epithelial cell lines (HIO 1120 and HIO 180). Tumor cells formed matrix-rich tubular networks containing channels surrounding spheroids of tumor cells, and this network may represent either a primitive microcirculatory-like network, or a remodeled vascularized portion of a tumor (Sood et al, 2001).