Morphological control has been achieved by increasing the inoculum size, changing the initial pH value of the medium, and using additives, such as carboxymethylcellulose (CMC), chelators, Tween-80, or agar granules.
Mammospheres were grown in serum-free, low adherence cultures, with a CnT-27 medium and growth additives (CellnTEC Advanced cell systems, Bern, Switzerland), as described by Dontu et al. [52].
In the process, the solvent medium and the surfactant additive CTAB played very important roles in the formation of different biomimetic morphologies and the formation mechanisms were primarily discussed, respectively.
Finally, Trypsin residues were removed with, first, centrifugation (1600 rpm/5 min) of the cell fluid (culture medium with additives and loose osteoblast cells) and, second, by subsequently adding 10 ml of culture medium.
The growth medium was then replaced by basal medium without additives and responses to isoproterenol, NNK and GABA assessed in untransfected cells versus cells with GABABR knockdown, using the cyclic adenosine monophosphate response element (CRE -luciferase reporter assay desribed below.
"One of the research projects we'll be undertaking is to use the local rock as a growing medium by adding sufficient minerals and additives".
Murashige and Skoog's (MS; Murashige and Skoog 1962) medium supplemented with sucrose (3 %) and additives (50 mg L−1 ascorbic acid, 25 mg L−1 adenine sulfate, 25 mg L−1 arginine and 25 mg L−1 citric acid), as described by Arya et al. (2002), was used for culture initiation.
In fact, the combination of low glucose in the culture medium and Dex had an additive effect on cell death in RS4 11 cells.
Culture medium, additives and antibiotics were obtained from Gibco BRL (Life Technologies, Paisley, UK) and Sigma Chemicals Ltd. (Poole, UK).
