Background: Schizophrenia is a strongly heritable multifactorial mental disorder whose aetiology is not fully understood. Lack of identified causative genes and the limitations of animal models have impeded the understanding of this disease from the biological perspective. There is a consensus, based on epidemiology and post-mortem human brains that schizophrenia is a neurodevelopmental disorder, but cellular and molecular mechanisms for this hypothesis are lacking. We have developed a model to address this using human olfactory neurosphere-derived (hONS) ‘stem’ cells, which allows comparative analyses between age- and sex-matched schizophrenic patients and healthy subjects. Patient cells have dysregulated focal adhesion dynamics and altered gene pathways for cell motility and adhesion. The aim of this work is to expand on what we previously know about migration/motility deficits in patient cells on fibronectin to other ECM matrices. This work also aims to develop an efficient, effective and quantitative cell motility assay.
Methodology: Cell cycle synchronised hONS cells will be seeded onto 96-well plate wells pre-coated with fibronectin, laminin, collagen as well as uncoated wells. Cells were allowed to attach for 4 h followed by addition of NucBlue® Live ReadyProbes™ Reagent (Life Technologies) to visualise cell nuclei. Time-lapse microscopy was used to track the coordinates of the stained nucleus over 24, 48 and 72 h at one frame every 10 min. Final time-lapse data will be analysed with Imaris (Bitplane). Dynamic measurements such as speed, displacement and distance travelled will be collated and compared between patient and control cells.