Title Page
Contents
ABSTRACT 6
I. INTRODUCTION 10
II. MATERIALS AND METHODS 12
1. Human placenta-derived mesenchymal stem cell (hPMSC) preparation and isolation of exosomes from hPMSCs 12
2. Cell culture and treatment 13
3. Small-interfering RNA 14
4. Cell viability assay 14
5. Reverse transcription polymerase chain reaction (RT-PCR) analysis 14
6. Immunoblot analysis 15
7. Proteomics 16
7.1. Materials 16
7.2. Sample preparation 17
7.3. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis 18
7.4. Data processing and bioinformatics 19
8. Statistical analyses 20
III. RESULTS 21
1. Characterization and recovery effects of hPMSC exosomes 21
2. Hierarchical clustering and gene ontology 22
3. Gene ontology classification of reliably quantified proteins from exosomes in hypoxia-damaged retinal precursor cells 24
4. Network analysis of recovery process mediators 26
5. Effects of hPMSC exosomes in hypoxia-damaged in vitro model 27
6. Role of UBA2 during recovery induced by hPMSC exosomes 29
IV. DISCUSSION 32
V. CONCLUSION 39
FIGURE LEGENDS 40
REFERENCES 43
Fig. 1. Characterization and recovery effects of hPMSC-derived exosomes 22
Fig. 2. Cluster analysis of differentially expressed proteins in R28 cells 23
Fig. 3. Gene ontology (GO) distribution of differentially expressed... 25
Fig. 4. Network analysis of DEPs relating to ubiquitin-mediated... 27
Fig. 5. Effect of exosome treatment on target proteins altered by CoCl₂ 29
Fig. 6. UBA2 mediation during recovery induced by exosomes 31
Fig. 7. Proposed pathway of recovery of R28 cells, induced by hPMSC-derived exosomes, after... 32