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To study the effect of paracrine mediators secreted by myoblast sheets, we washed the sheets thoroughly with serum free medium, incubated the sheets in that medium for 24 hours, and collected the conditioned medium for experiments.
When adhered, cells were switched to AIM-V serum free medium for experiments.
The following day, the transfection mixture was removed and the appropriate medium for experiments was added.
Dialysed serum was used in the cell growth medium for experiments involving 5HT2c ligands.
5-Fluorouracil (5-FU; Wako Pure Chemical Industries, Ltd, Osaka, Japan) and CDDP (Wako) were dissolved in dimethylsulphoxide and then diluted in culture medium for experiments.
After sonication, stock solutions were mix with a vortex for 1 min, and various concentrations of SPION were prepared in culture medium for experiments.
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The minimal medium for experiment cultures contained a mixture of 20% [U-C] labeled glucose (C enrichment ≥ 99%, Cambridge Isotope Laboratories, Andover, USA) and 80% naturally labeled glucose.
The growth medium used for experiments is also very important because genes essentiality is dependent on what substrates are available for use.
False negative errors can be caused by incomplete definition of the biomass function, uncertainty in the growth medium used for experiments, and toxic-intermediate buildup.
This underlines the importance of measuring the actual agglomerate sizes of NPs in the cell culture medium used for experiments, rather than relying on zeta potential measurements to predict likely agglomeration behaviour.
In order to induce increased stress, ARPE-19 cells were placed directly after splitting into serum-free medium for all experiments and incubated for up to 72 h in a humidified cell incubator at +37 °C.
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