Transient maintenance in bioreactor improves health of neuronal cells
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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 inter-leukin-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.
Key wordsaggregated cultures neurons apoptosis AOE PPARs cytokines
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- Aebi, H. E. Catalase. In: Bergmayer, H., ed. Methods in enzymatic analysis, 2nd ed. Wenheim: Verlag Chemic; 1974:673–684.Google Scholar
- Bonfoco, E.; Kraine, D.; Ankarcrona, M.; Nicotera, P.; Lipton, S. A. Apoptosis and necrosis: two distinct events induced, respectively, by mild and intense insults with N-methyl-d-aspartate or nitric oxide/superoxide in cortical cell cultures. Proc. Natl. Acad. Sci. USA 92:7162–7166; 1995.PubMedCrossRefGoogle Scholar
- Gruener, R.; Hoeger, G. Vector-free gravity disrupts synapse formation in cell culture. Am. J. Physiol. 258:489–494; 1990.Google Scholar
- Kitamura, Y.; Shimohama, S.; Koike, H.; Kakimura, J.; Matsuoka, Y.; Nomura, Y.; Gebike-Haerter, P. J.; Taniguchi, T. Increased expression of cyclooxygenase and peroxisome proliferator-activated receptor-γ in Alzheimer's disease brains. Biochem. Biophys. Res. Commun. 254:582–586; 1999.PubMedCrossRefGoogle Scholar
- Lelkes, P. I.; Galvan, D. L.; Hayman, G. T.; Goodwin, T. J.; Chatman, D. Y.; Cherian, S.; Garcia, R. M.; Unsworth, B. R. Simulated microgravity conditions enhance differentiation of cultured PC12 cells towards the neuroendocrine phenotype. In Vitro Cell. Dev. Biol. 34A:316–325; 1998.Google Scholar
- Medina, J.; Tabernero, A. Astrocyte-synthesized oleic acid behaves as neurotrophic factor for neurons. J. Physiol. 96:265–271; 2002.Google Scholar
- Yamada, M.; Iwatsubo, T.; Mizuno, Y.; Mochizuki, H. Overexpression of alpha-synuclein in rat substantia nigra results in loss of dopaminergic neurons, phosphorylation of alpha-synuclein and activation of caspase-9: resemblance to pathogenetic changes in Parkinson's disease. J. Neurochem. 91:451–461; 2004.PubMedCrossRefGoogle Scholar