The Use of Adipose Derived Stem Cells in Spinal Cord and Peripheral Nerve Repair the Use of Adipose Derived Stem Cells in Spinal Cord and Peripheral Nerve Repair Table of Contents

6 ORIGINAL PAPERS 7 ABBREVIATIONS 8 INTRODUCTION 1. Clinical background and epidemiology 9 2. Pathophysiology of spinal cord and peripheral nerve injury 2.1. Spinal cord 9 2.1.1. Acute and sub-acute spinal cord injury 10 2.1.2. Chronic spinal cord injury 11 2.2. Peripheral nerve 11 3. Regeneration following spinal cord and peripheral nerve injury 3.1. Regeneration following spinal cord injury 12 3.2. Regeneration following peripheral nerve injury 12 3.3. Biosynthetic conduits for peripheral nerve repair 13 4. Cell transplantation to promote spinal cord and peripheral nerve repair 14 AIMS OF THE STUDY 15 MATERIALS AND METHODS 1. Cell culture 1.1. Culture of human adipose derived stem cells 16 1.2. Culture of dorsal root ganglion neurons and neurite outgrowth assay 17 1.3. Culture of cortical astrocytes 17 1.4. Culture of Schwann cells 17 1.5. Confrontation assay of astrocytes with ASC and Schwann cells 18 1.6. In vitro angiogenesis assay 18 2. Experimental animals and surgery 18 2.1. Peripheral nerve injury 19 2.2. Spinal cord injury 19 3. Tissue processing and analysis 3.1. Tissue harvest 20 3.2. RT-PCR 20 3.3. qRT-PCR 21 3.4. Western blotting 21 3.5. Enzyme-linked immunosorbant assay (ELISA) 22 3.6. Immunostaining 22 3.7. Analysis of regeneration in fibrin conduits 22 3.8. Analysis of regeneration in spinal cord 23 4. Image processing 23 5. Vertical cylinder exploration test 23 6. Statistical analysis 23