Brief Article
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World J Stem Cells. Oct 26, 2013; 5(4): 229-237
Published online Oct 26, 2013. doi: 10.4252/wjsc.v5.i4.229
Neural stem cells isolated from amyloid precursor protein-mutated mice for drug discovery
Vito Antonio Baldassarro, Giulia Lizzo, Michela Paradisi, Mercedes Fernández, Luciana Giardino, Laura Calzà
Vito Antonio Baldassarro, Giulia Lizzo, Mercedez Fernández, Health Science and Technologies Interdepartmental Center for Industrial Research (HST-ICIR), I-40064 Ozzano Emilia, Bologna, Italy
Michela Paradisi, Department of Veterinary Medicine, University of Bologna, I-40064 Ozzano Emilia, Bologna, Italy
Luciana Giardino, Laura Calzà, Health Science and Technologies Interdepartmental Centre for Industrial Research (HST-ICIR) and IRET Foundation, I-40064 Ozzano Emilia, Bologna, Italy
Giulia Lizzo, Institut de Recherches Servier, 125 Chemin de Ronde, 78290 Croissy-Sur-Seine, Yvelines, France
Author contributions: Baldassarro VA wrote the manuscript and performed part of the experiments; Lizzo G and Paradisi M performed the majority of the experiments; Fernandez M set up the techniques and contributed to the manuscript; Giardino L and Calzà L designed the study and revised the manuscript.
Supported by Regione Emilia Romagna, POR-FESR 2011-2014
Correspondence to: Laura Calzà, MD, Health Science and Technologies Interdepartmental Centre for Industrial Research (HST-ICIR), University of Bologna, Via Tolara di Sopra 41/E, I-40064 Ozzano Emilia, Bologna, Italy. laura.calza@unibo.it
Telephone: +39-51-798776 Fax: +39-51-799673
Received: June 24, 2013
Revised: October 4, 2013
Accepted: October 17, 2013
Published online: October 26, 2013
Processing time: 128 Days and 18.1 Hours
Abstract

AIM: 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.

CONCLUSION: Tg2576 NSCs represent an appropriate AD in vitro model resembling some cellular alterations observed in vivo, both as stem and differentiated cells.

Keywords: Neural stem cells; Alzheimer’s disease; Neuron maturation; Drug discovery

Core tip: In this study neural stem cells isolated from Tg2576 mice are characterized as an in vitro model for Alzheimer’s disease. These cells represent a robust system for studying pathological mechanisms related to Aβ intracellular accumulation, such as stem cell status, or during differentiation processes. This model could provide a new cell platform for developing and screening new molecules.