Gentamicin Sulfate, USP is an aminoglycoside antibiotic complex discovered in 1963 derived from fermentation of Micromonospora purpurea or M. echinospora. Gentamicin is composed of different components including Gentamicin C complex (gentamicin C1, gentamicin C1a, and gentamicin C2) which makes up 80% of the compound and has the highest antibacterial activity, along with Gentamicin A, B, X, and a few others which make up the remaining 20% of Gentamicin and have lower antibiotic activity. Gentamicin Sulfate is suitable for use in cell culture to prevent and control bacterial contamination and the compound is soluble in water (50 mg/ml).
Gentamicin Sulfate, USP conforms to United States Pharmacopoeia specifications.
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|Mechanism of Action||Aminoglycosides are a widespread and versatile group of bioactive natural products. They target the 30S ribosomal subunit, blocking the translocation of peptidyl-tRNA from acceptor to donor. This results in an inability to read mRNA ultimately producing a faulty or nonexistent protein.|
|Spectrum||Gentamicin Sulfate 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."|
|Impurity Profile||Gentamicin C1: 25.0 -50.0% |
Gentamicin C1a: 10.0 - 35.0%
Gentamicin C2a + C2: 25.0 - 55.0%
|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. |
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 inhibited differentiation of tracheary elements in pith parenchyma cells in cultures of romaine lettuce (Lactuca sativa L. var. Romana) at concentrations of 50-100 µg/ml. Similar results were obtained with cultured explants of Jerusalem artichoke tuber (Helianthus tuberosus L.). Callus formation was suppressed with increasing levels of Gentamicin Sulfate in both tissue systems. When studying cell division or xylem differentiation in culture, it is best to use ≤ 10 µg/ml.|
|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).
Porcine intestinal epithelial cell line IPEC-J2 cell monolayers were used to study the toxicity of Gentamicin in cell culture. Authors found it did not affect IPEC-J2 cell monolayer integrity via the disruption of cell membranes and did not increase the incidence of toxic effects related to membrane permeation (Gyetvai et al, 2015).
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).|
Bürgi N, Josi C, Bürki S and Schweizer, Pilo P (2018) Mycoplasma bovis co-infection with bovine viral diarrhea virus in bovine macrophages. Vet. Res. 49(1):2. PMID 29316971Gyetvai B et al (2015) Gentamicin sulphate permeation through porcine intestinal epithelial cell monolayer. Act. Vet. Hung. 63(1): 60-68 PMID 25655415
Kadurugamuwa JL, Clarke AJ and Beveridge TJ (1993) Surface action of gentamicin on Pseudomonas aeruginosa. J. Bacteriol 175(18):5798-5805 PMID 8376327
Martin NL and Bevridge TJ (1986) Gentamicin interaction with Pseudomonas aeruginosa. Antimicrob. Agents Chemother. 29(6):1079-1087 PMID 2425732
Montenez JP, Kishore BK, Maldaque P and Tulkens PM (1984) Leupeptin and E-64, inhibitors of cysteine proteinases, prevent gentamicin-induced lysosomal phospholipidosis in cultured rat fibroblasts. Toxicol Lett. 73(3):201-208 PMID 8091428
Rudin A, Healey A, Phillips CA, Gump DW and Forsyth BR (1970) Antibacterial activity of gentamicin sulfate in tissue culture. Appl. Microbiol. 20(6):989-990. PMID 4992660
Sood AK (2001) Molecular determinants of ovarian cancer plasticity. Am. J. Pathol. 158(4):1279-1288. PMID 11290546
Temel Y, Ayna A, Shafeeq IH and Ciftci M (2018) In vitro effects of some antibiotics on glucose-6-phosphate dehydrogenase from rat (Rattus norvegicus) erythrocyte. Drug and Chemical Toxicol. DOI: 10.1080/01480545.2018.1481083
Wan J et al (1994) Intravesical instillation of gentamicin sulfate: In vitro, rat, canine, and human studies. Urology 43(4):531-536. PMID 8154077
|MIC||Bacillus cereus| 1.22|| Bacillus subtilis| 0.5|| Escherichia coli| 1.22 - 2.44 || Klebsiella pneumonia| 1|| Pseudomonas aeruginosa| 0.5 - 2.44 || Salmonella enterica| 1|| Staphylococcus aureus| 0.5 - 0.61 || Stenotrophomonas maltophilia| 2 - >16 |||