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
Articles Cell Growth/Differentiation/Apoptosis

Summary

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 words

aggregated cultures neurons apoptosis AOE PPARs cytokines 

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Copyright information

© Society for In Vitro Biology 2006

Authors and Affiliations

  • Silvia Di Loreto
    • 2
    • 1
  • Pierluigi Sebastiani
    • 2
  • Elisabetta Benedetti
    • 1
  • Vincenzo Zimmitti
    • 1
  • Valentina Caracciolo
    • 1
  • Fernanda Amicarelli
    • 1
  • Annamaria Cimini
    • 1
  • Domenico Adorno
    • 2
  1. 1.Department of Basic and Applied BiologyUniversity of L'AquilaL'AquilaItaly
  2. 2.Institute of Organ Transplants and ImmunocytologyConsiglio Nazionale delle RicercheL'AquilaItaly

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