Patients with multiple system atrophy (MSA), a progressive neurodegenerative disorder of adult onset, were found less than 9-year life expectancy after onset. The disorders include bradykinesia and rigidity commonly seen in Parkinsonism disease and additional signs such as autonomic dysfunction, ataxia, or dementia. MSA-P, representing 80% of MSA, is characterized by striatonigral neurodegeneration (SND) where parkinsonism predominates. In clinical treatments, MSA poorly responds to levodopa, the drug used to remedy Parkinsonism disease. The exact cause of MSA is still unknown and exploring a therapeutic solution to MSA remains critical. The potential of stem cell therapy for neural degenerative diseases has been widely explored recently. With the reported abilities to differentiate into neuronal linage and to secrete neurotrophic factors for neuronal support, mesenchymal stem cells (MSC) presents a possible therapeutic option for MSA-P. Among the sources of MSC, adipose-derived stem cells (ADSC) are relatively easier to obtain and less invasive. To conduct the feasibility study in vivo, toxin-based MSA-P mouse model and transgenic MSA mouse model were established. Toxin-based model, chemically (3NP + MPTP) inducing MSA-P in C57BL/6N mice, was established to assess the effect of ADSC on improving motion behavior of MSA-P mouse. Molecular mechanism was investigated using transgenic MSA mouse model to reveal the role of ADSC in alleviating neurodegeneration of MBP1 mouse. MBP1 transgenic mice, transferred from the University of California in San Diego, USA (UCSD), have been reported in studying α-synuclein associated neurodegeneration. Our transgenic mouse model established from MBP1 mouse provides a useful tool to investigate the effect of ADSC on the biomarker for MSA, i.e., α-synuclein glial cytoplasmic inclusion (GCI) in oligodendrocytes. Human ADSC were transplanted into the striatum of MSA animals via intracerebral injection (IC). Testing intranasal (INA) delivery route was withdrawn due to brain blood barrier (BBB) issue. The grafted ADSC was traced by labeling ADSC with transduced GFP and taking photo images on live mice. The presence of ADSC was detected from immunofluorescence (IF) stains 4 weeks post grafting. The alleviation of neurodegeneration after ADSC treatment was indicated by improved performance in animal behavior testing, including rotarod test and pole test. As compared with a sham control, ADSC significantly enhanced Rotarod performance of MSA animals treated with ADSC at an effective dose (2x10^5 ADSC/mouse). Our ex vivo study supported that ADSC might alleviate striatal degeneration in MSA animal model by improving nigrostriatal pathway for dopamine, activating autophagy for α-synuclein clearance, decreasing inflammatory signal and further cell apoptosis, improving myelination and cell survival at caudate-putamen. Mitochondria was found to transfer from ADSC to injured neuron in vitro. ADSC pretreated with mitochondria-rich chemical (MR1) had beneficial trend in improving Rotarod performance although the difference against ADSC was not statistically significant.

Chapter I INTRODUCTION 1
Chapter II MATERIALS AND METHODS 7
II-A. Preparing ADSC for feasibility study 7
II-B. Establishing chemically induced mouse model and investigating the effect of ADSC 8
II-C. Establishing SHSY5Y cell model to track mitochondria of ADSC, in vitro 12
II-D. Establishing α-synuclein over-expressed transgenic mouse model and investigating the effect of ADSC 13
II-E. Studying the mechanism using the α-synuclein over-expressed transgenic mouse model 15
Chapter III RESULTS 19
III-A. Preliminary Findings from Chemically Induced Models Supported Therapeutic Potential of ADSC for Treating MSA-P Disease 19
III-B. MSA Symptoms Revealed by the Over-expression of α-synuclein Driven by MBP Promotor 20
III-C. Beneficial Effects on the MSA Transgenic Mouse Model Revealed by ADSC Transplantation 20
III-D. Nigrostriatal Pathway for Dopamine Signal Improved by ADSC Transplantation 21
III-E. Potential of ADSC Transplantation in Activating Autophagy Protein Clearance Pathway to Decrease the Levels of Un-aggregated Form of α-synuclein 22
III-F. Potential of ADSC Transplantation in Decreasing Inflammatory Signal and Further Cell Apoptosis 23
Chapter IV DISCUSSION 25
Chapter V CONCLUSIONS 31
Chapter VI FUTURE WORK 33
REFERENCES 35
TABLES AND FIGURES 49

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