The human embryonic kidney 293 cell line (HEK 293, ECACC 85120602) was provided by Sigma-Aldrich (Saint-Quentin Fallavier, France). The hepatoma cell line HepG2 was provided by ECACC (85011430). JEG3 cell line (ECACC 92120308) was provided by CERDIC (Sophia-Antipolis, France). Cells were grown in phenol red-free EMEM (Abcys, Paris, France) containing 2 mM glutamine, 1% nonessential amino acid, 100 U/mL of antibiotics (a mixture of penicillin, streptomycin, and fungizone) (Lonza, Saint Beauzire, France), 10 mg/mL of liquid kanamycin (Dominique Dutscher, Brumath, France), and 10% Fetal Bovine Serum (PAA, les Mureaux, France). JEG3 cells were supplemented with 1 mM sodium pyruvate. Cells were grown with this medium at 37°C (5% CO2, 95% air) during 48 h to 80% confluence, then washed, and exposed 24 h with serum-free EMEM to the APs or the formulations. Before treatment, all the pesticides were solubilized in a 100% DMSO solution, then diluted in serum-free medium to reach 0.5% DMSO (which had been previously proven not to be cytotoxic for the cells), and adjusted to a similar pH. This model was validated [29 (link)] and cytotoxic effects were similar in presence of serum but delayed by 48 h.
Liquid kanamycin
Manufactured by Dutscher
Sourced in France
Liquid kanamycin is an antibiotic solution commonly used in microbiology and molecular biology applications. It provides a source of the antibiotic compound kanamycin, which is used to select for bacterial cells that have been successfully transformed or transfected with genetic material.
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2 protocols using liquid kanamycin
1
Standardized in vitro cell culture protocol
2
Cytotoxicity Evaluation of Glyphosate-Based Herbicides
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The human embryonic kidney 293 cell line (HEK 293, ECACC 85120602) was provided by Sigma-Aldrich (Saint-Quentin Fallavier, France). JEG3 cell line (ECACC 92120308) was provided by CERDIC (Sophia-Antipolis, France). Both were validated for toxicity studies of pesticides [8 (link)], corresponding to what is observed for fresh tissue or primary cells [7 (link)]. These cell lines are even in some instances less sensitive than primary cells [9 (link)], and therefore do not overestimate cellular toxicity. Cells were grown with log-tested methods in phenol red-free EMEM (Abcys, Paris, France) containing 2 mM glutamine, 1% non-essential amino acid, 100 U/mL of antibiotics (a mixture of penicillin, streptomycin and fungizone) (Lonza, Saint Beauzire, France), 10 mg/mL of liquid kanamycin (Dominique Dutscher, Brumath, France) and 10% Fetal Bovine Serum (PAA, les Mureaux, France). JEG3 cells were supplemented with 1 mM sodium pyruvate. Cells were grown in this medium at 37 °C (5% CO2, 95% air) during 48 h to 80% confluence, then washed and exposed for 24 h with serum-free EMEM to the formulations of GBH, formulants and G or its salt, diluted in serum-free medium and adjusted to a similar pH. This model has been carefully validated [10 (link)] since cytotoxic effects were similar in the presence of serum but delayed by 48 h, and some compounds of the serum may interfere with the test.
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