Transient maintenance in bioreactor improves health of neuronal cells.

To examine whether a neuronal cell suspension can be held in vitro for a relatively short period without compromising
survival rates and functionality, we have set up an experimental protocol planning 24 h of suspension culture in a rotary
wall vessel (RWV) bioreactor before plating in a conventional adherent system. Apoptosis measurement and activated
caspase-8, -9, and -3 detection have demonstrated that survey of the cells was not affected. The activity of major
antioxidant enzymes (AOE), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT), was
significantly decreased in RWV-maintained cells. A significant decrease of tumor necrosis factor- (TNF-) and interleukin-
1 (IL-1) is coupled with a level of activated nuclear factor-B (NF-B) protein significantly lower in RVW cells
than in the control. On the contrary, the level of IL-6 expression did not change between the test and the control. A
significant up-regulation of growth-associated protein-43 (GAP-43), peroxisome proliferator-activated receptor-/
(PPAR/), and acyl-CoA synthetase 2 (ACS2) in RWV cells has been detected. We provide the evidence that primary
neuronal cells, at an early stage of development, can be maintained in a suspension condition before adherent plating.
This experimental environment does not induce detrimental effects but may have an activator role, leading cells to
development and maturation in a tridimensional state.