Transient Maintenance in Bioreactor Improves Health of Neuronal Cells.

Bioreactor Transient Exposure Activates Specific Neurotrophic Pathway in Cortical Neurons

“…On the contrary, bioreactor transient exposition induces long-term growth associated proteins expression (Di Loreto et al 2006), leading neuronal cells to tri-dimensional aggregation feature, with elongated axons that connect the clusters, mimicking the neuronal network in a typical grey/white-matter fashion observed in vivo. It has been widely demonstrated that neurotrophins, specifically NGF and BDNF, have an important role in neural activity-dependent development and plasticity of brain and particularly of cortex.…”

“…Previously, utilizing a model of transient maintenance in RWV, we had demonstrated, with morphological and electro-functional evidences, that transient cultivation in RWV does not significantly affect the morphological and electrophysiological development and maturation of neuronal cell in a followup adherent culture (Crestini et al 2004). Recently, we have extended these findings investigating the apoptotic, oxidative mechanisms, proinflammatory cytokine profile and growth-related proteins (Di Loreto et al 2006). We have demonstrated that this experimental environment not only does not induce detrimental effects but the cultures maintained in the RWV system for 24 h showed biomolecular parameters suggesting the activation of a pathway that leads neural cells to survive better and to develop in a tri-dimensional state.…”

“…The RWV bioreactor (Synthecon, Houston, TX) was designed to grow cell cultures into a three-dimensional mass and the procedure and devices were previously described (Crestini et al 2004;Di Loreto et al 2006). The cell density in the bioreactors was approximately 1 × 10 6 cells/ml.…”

“…All procedures were carried out as previously described (Di Loreto et al 2006). Briefly, total cellular RNA was extracted by Trizol Reagent (Invitrogen) according the manufacturer's instructions and retro-transcripted to cDNA using random decamers as first-strand primers.…”

“…Previous work widely described the western blot procedures (Di Loreto et al 2006). Briefly, cells were lysed in RIPA solution and lysates were cleared by centrifugation at 12,000 rpm for 20 min.…”