One millilitre of C2C12 cells at a concentration of 5 x 104 cells/mL were seeded in a 24-well culture plate and attached overnight in Dulbecco's modified Eagle's medium (DMEM) with 1% (v/v) fetal calf serum (FCS), 1% L-glutamine and no antibiotics at 37°C, 5% CO2, in a humidified environment.
Alternatively, passage II IVD cells were seeded into alginate beads at a low cell density of 106 cells per millilitre and a final alginate concentration of 1.2%.
One millilitre containing 1.2 × 10 cells was seeded into each well of a 24-well plate coated with collagen IV (BD Biosciences, Bedford, MA, USA).
The average radioactivity concentrations in the ROIs (kilobecquerels per millilitre) were used for further analyses.
CFU per millilitre were counted after incubation for 2 days at 30 °C.
The change in sperm concentration to 49.9m per millilitre is still within the normal range, and above the lower threshold of 15m per millilitre used by fertility doctors, said Pacey.
The number of Salmonella bacteria per millilitre was determined by plating each dilution onto TSA plates.
The mean concentration of viable bacteria per millilitre is indicated with error bars representing standard error.
The results, published on Wednesday in the journal Biology Letters, showed that the biggest brains, averaging 1,484 millilitres, were from Scandinavia, while the smallest brains, around 1,200 millilitres, came from Micronesia.
Typical optical absorption spectra of gold nanostructures prepared using several concentration ratios of CTAB and HMT (millilitre to millilitre ratio) are shown in Figure 4.
