title: Neural stem cells isolated from amyloid precursor protein-mutated mice for drug discovery.
abstract: OBJECTIVE
  To develop an in vitro model based on neural stem cells derived from transgenic animals, to be used in the study of pathological mechanisms of Alzheimer's disease and for testing new molecules.
  
  
  METHODS
  Neural stem cells (NSCs) were isolated from the subventricular zone of Wild type (Wt) and Tg2576 mice. Primary and secondary neurosphere generation was studied, analysing population doubling and the cell yield per animal. Secondary neurospheres were dissociated and plated on MCM Gel Cultrex 2D and after 6 d in vitro (DIVs) in mitogen withdrawal conditions, spontaneous differentiation was studied using specific neural markers (MAP2 and TuJ-1 for neurons, GFAP for astroglial cells and CNPase for oligodendrocytes). Gene expression pathways were analysed in secondary neurospheres, using the QIAGEN PCR array for neurogenesis, comparing the Tg2576 derived cell expression with the Wt cells. Proteins encoded by the altered genes were clustered using STRING web software.
  
  
  RESULTS
  As revealed by 6E10 positive staining, all Tg2576 derived cells retain the expression of the human transgenic Amyloid Precursor Protein. Tg2576 derived primary neurospheres show a decrease in population doubling. Morphological analysis of differentiated NSCs reveals a decrease in MAP2- and an increase in GFAP-positive cells in Tg2576 derived cells. Analysing the branching of TuJ-1 positive cells, a clear decrease in neurite number and length is observed in Tg2576 cells. The gene expression neurogenesis pathway revealed 11 altered genes in Tg2576 NSCs compared to Wt.
  
  
  CONCLUSIONS
  Tg2576 NSCs represent an appropriate AD in vitro model resembling some cellular alterations observed in vivo, both as stem and differentiated cells.
authors.affiliation: Vito Antonio Baldassarro, Mercedez Fernández, Giulia Lizzo, Health Science and Technologies Interdepartmental Center for Industrial Research (HST-ICIR), I-40064 Ozzano Emilia, Bologna, Italy.
authors.forename: Vito Antonio
authors.initials: VA
authors.lastname: Baldassarro
authors.forename: Giulia
authors.initials: G
authors.lastname: Lizzo
authors.forename: Michela
authors.initials: M
authors.lastname: Paradisi
authors.forename: Mercedes
authors.initials: M
authors.lastname: Fernández
authors.forename: Luciana
authors.initials: L
authors.lastname: Giardino
authors.forename: Laura
authors.initials: L
authors.lastname: Calzà
keywords: Alzheimer’s disease; Drug discovery; Neural stem cells; Neuron maturation