| Parkinson's disease (PD) is a chronic and progressive neurological disorder resulting from the loss of dopaminergic neurons of the substantia nigra of the brain. A three-dimensional (3D) culture model that better represents the cytoarchitecture of the in vivo environment could provide a more accurate research tool than animal or 2D models, possibly leading to earlier diagnosis and better outcomes for patients. The model could be used to ethically and safely test novel treatment methods, such as stem cell therapies, and provide a template for the development of implantable therapies for injured or diseased tissues or cells. PC12 cells seeded within a 3D collagen gel matrix survive and proliferate in the undifferentiated state for up to nine days in culture, and respond to 100ng/ml nerve growth factor by ceasing proliferation and extending neuritic processes. Differentiated cells express tyrosine hydroxylase, an indicator of dopaminergic neurons, and maintain their differentiated state for up to nine days in the 3D cell culture.; Bone marrow stromal cells (BMSC) have great potential as cell-based therapies for the treatment of neurodegenerative disorders: they are pluripotent, are relatively easily harvested, proliferate well in culture, and have the potential of being harvested from a patient's own body, thereby circumventing immune rejection concerns. Electromagnetic fields (EMF) have been shown to affect the dendritic outgrowth and orientation of neuronal cells in vitro , and to induce axonal outgrowth in vivo. A multi-factor induction protocol causes neuronal differentiation of BMSC into specific neural cell types, resulting in significantly higher numbers of differentiated cells as compared to single-factor protocols (p < 0.005). A collagen fiber/collagen gel 3D matrix is a viable environment for both differentiated and undifferentiated BMSC. Exposing differentiated BMSC in 2D cultures to an EMF-induced electric field of 1--5V/cm affects the orientation of neuronally differentiated BMSC in vitro. PC12 cells seeded within a collagen gel matrix and differentiated through exposure to NGF are not significantly affected after 7 days' exposure to 30 minutes per day of a 10Hz, 2V/cm EMF. |
