Title

Developing a novel co-culture model and testing the effects of placental derived stem cells on inflammation in Alzheimer's disease.

Date of Award

5-1-2014

Document Type

Thesis

School

College of Liberal Arts

Degree Name

Bachelor in Arts

Abstract

Alzheimer's disease (AD) is an abhorrent disease that robs people of their memories and personalities. Researchers have yet to develop an effective disease-modifying treatment for this multifaceted disease. Inflammation is one of the main hallmarks of AD and is caused by the over-activation of the main immune cells of the brain: neuroglial cells. High concentrations of nitric oxide (NO) associated with this heightened immune response have been known to cause neuronal damage and cellular loss. The aim of this study was to develop a viable co-culture system in which placental derived stem cells (PDSCs) and neuroglial cells are cultured together in systems involving transwells and cover slips for further analysis. A transwell co-culture system resulted in no effect of PDSCs on the NO concentration mediated by LPS stimulation or the modulation of NGF in glia cultures. This suggests that NGF may not necessarily be the neurotrophin causing the decrease of NO in activated glia. A second co-culture system, a cell-to-cell contact cover slip system in which glia were grown in a welled plate and PDSCs grown on cover slips, revealed a decrease in NO concentration in LPS stimulated glia when compared to controls. There were several problems with this model however, including cell disruption stemming from execution flaws. Using Immunocytochemistry, this study also qualified and quantified the morphological differences in activated glial cells that had been treated with stem cells visualized through an induced nitric oxide synthase antibody (iNOS). LPS stimulated glia showed a shrunken, shriveled appearance typical of apoptosis with a high intensity of iNOS stain and a high area percentage covered by stain. Control glia, however, showed healthy spread and flattened glial formations with a minor but widespread intensity of iNOS. These studies, though preliminary, offer a platform on which to conduct further experiments in glia and PDSC co-culture as well as clues about how iNOS gets activated in glial inflammation in a model of AD.