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-α (TNF-α) and interleukin-1β (IL-1β) is coupled with a level of activated nuclear factor-κB (NF-κB) 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-β/δ (PPARβ/δ), 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.
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1 May 2006
TRANSIENT MAINTENANCE IN BIOREACTOR IMPROVES HEALTH OF NEURONAL CELLS
SILVIA DI LORETO,
PIERLUIGI SEBASTIANI,
ELISABETTA BENEDETTI,
VINCENZO ZIMMITTI,
VALENTINA CARACCIOLO,
FERNANDA AMICARELLI,
ANNAMARIA CIMINI,
DOMENICO ADORNO
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In Vitro Cellular & Developmental Biology - Animal
Vol. 42 • No. 5
May 2006
Vol. 42 • No. 5
May 2006
aggregated cultures
AOE
Apoptosis
cytokines
neurons
PPARs