The neuroblastoma cells were cultured in Dulbecco's modified eagle medium containing nutrient mixture F-12 (Gibco, Thermo Fisher Scientific Inc., Waltham, MA, USA) with 10% fetal bovine serum (Gibco, USA) and 1% penicillin-streptomycin-neomycin antibiotic mixture (Gibco, USA) and maintained at 37°C in 5% CO2 humidified air.
Fifty (50) mL of inoculum medium containing nutrient broth 13 g/L, pH 7.4, was transferred to a 250 mL Erlenmeyer flask and was sterilized in an autoclave (CL-40M, Japan) at 15 lbs/inch pressure at 121°C for 20 min.
By contrast, 4 h incubation with niclosamide, rottlerin and perhexiline at concentrations that were sufficient to profoundly inhibit mTORC1 signaling and stimulate autophagy had little or no effect on cell viability or proliferation in cell culture medium containing nutrients and serum.
We found that culturing retinas from ND embryos ex vivo in L15 medium, containing nutrients but no serum or growth factors, resulted in a significant increase in CMZ proliferation (Fig. 4A,B), demonstrating functionally that ND CMZ cells are not lost and are still responsive to growth-promoting conditions.
A complex medium contains nutrients which are not well defined chemically e.g., yeast extract, tryptone, peptone, casamino acids, while defined medium consists of chemicals which are chemically defined.
The medium contained nutrients and organic carbon and samples were incubated under anaerobic conditions with pH of 7.5, in order to stimulate the growth of heterotrophic and anaerobic bacteria from the soil.
Then, the microalgal strain was grown on a petri plates consisting of agar medium containing the nutrient components (Tables 1 and 2) and agar (12.0 g/L).
EVLC2 cells were maintained in Dulbecco's Modified Eagle Medium containing F12 nutrient mixture, L-glutamine and pyruvate (DMEM; Invitrogen, Paisley, UK) and supplemented 10% foetal bovine serum (FBS; Promega, Southampton, UK).
Solidified nutrient agar was cast onto petriplates, and the plates containing nutrient medium were evenly inoculated with 100 μg (108 CFU/ml) separately.
Germ buds were transferred to plastic boxes containing nutrient medium for 10 weeks.
One milliliter of appropriate dilution suspension was transferred into Petri plates containing nutrient agar medium and then those plates were incubated at 37 °C for 24 h.
