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-alpha (TNF-alpha) and interleukin-1beta (IL-1beta) is coupled with a level of activated nuclear factor-kappaB (NF-kappaB) 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-beta/delta (PPARbeta/delta), 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.
Di Loreto
Silvia
S
Institute of Organ Transplants and Immunocytology, Consiglio Nazionale delle Ricerche, L'Aquila, Italy. sdiloreto@itoi.cnr.it
Sebastiani
Pierluigi
P
Benedetti
Elisabetta
E
Zimmitti
Vincenzo
V
Caracciolo
Valentina
V
Amicarelli
Fernanda
F
Cimini
Annamaria
A
Adorno
Domenico
D
D000818:Animals; D000975:Antioxidants / Q000378:metabolism; D017209:Apoptosis; D019149:Bioreactors; D002374:Catalase / Q000378:metabolism; D018929:Cell Culture Techniques / Q000295:instrumentation*; D003066:Coenzyme A Ligases / Q000235:genetics / Q000378:metabolism; D019922:GAP-43 Protein / Q000235:genetics / Q000378:metabolism; D005786:Gene Expression Regulation; D005979:Glutathione Peroxidase / Q000378:metabolism; D007375:Interleukin-1 / Q000235:genetics / Q000378:metabolism; D015850:Interleukin-6 / Q000235:genetics / Q000378:metabolism; D009474:Neurons / Q000166:cytology*; D047494:PPAR delta / Q000235:genetics / Q000378:metabolism; D048068:PPAR-beta / Q000235:genetics / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D013482:Superoxide Dismutase / Q000378:metabolism; D014409:Tumor Necrosis Factor-alpha / Q000235:genetics / Q000378:metabolism
D000975:Antioxidants; D019922:GAP-43 Protein; D007375:Interleukin-1; D015850:Interleukin-6; D047494:PPAR delta; D048068:PPAR-beta; D014409:Tumor Necrosis Factor-alpha; D002374:Catalase; D005979:Glutathione Peroxidase; D013482:Superoxide Dismutase; C500899:Acsl6 protein, rat; D003066:Coenzyme A Ligases