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All media was purchased from the media preparation facility at MSKCC (New York, NY).
Tissue culture procedures and culture media preparation have been previously described [5].
Agar (16 g/L) and/or Nile-red (final concentration 0.5 µg/mL), was added for solid media preparation.
Samples and media preparation, inoculation and colony counts were done using standard microbiological methods (Buckle et al. 1989; Bartram and Balance 1996; FAO 2004).
The Cu2+ concentration that the nano-CuO released to the culture media was measured by ICP-MS 7500ce, Agilent, USA at 24 h after media preparation.
Phosphate buffered saline (PBS) and Dulbecco's Modified Eagle Medium (DMEM) were purchased from the media preparation facility at Memorial Sloan Kettering Cancer Center (New York, NY, USA).
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A first study of the comparison of structures of enzymes (by FT-IR and CD) in different high activity (in low water media) preparations is reported.
Both media preparations hardly supported biofilm formation.
Media preparations containing NGF and BDNF were replenished every 3 DIV for CHOLNs and MSNs, respectively.
In any case, the results are indicative of ongoing enzymatic activity during storage of CMp11 and suggest snap freezing and subsequent storage at −80°C, or the use of fresh media, would likely minimise variance between media preparations.
All media preparations were autoclaved for 15 min.
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