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척수재생을 위한 신경줄기세포와 고분자 지지체를 이용한 복합적인 치료전략의 개발
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 김 병곤 | - |
| dc.contributor.author | 강영미 | - |
| dc.date.accessioned | 2019-10-21T07:11:28Z | - |
| dc.date.available | 2019-10-21T07:11:28Z | - |
| dc.date.issued | 2009-02 | - |
| dc.identifier.other | 9802 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/17069 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :의생명학과,2009. 2 | - |
| dc.description.tableofcontents | TABLE OF CONTENTS ABSTRACT i TABLE OF CONTENTS iii LIST OF FIGURES v LIST OF ABBREVIATION vi I. INTRODUCTION 1 II. MATERIALS AND METHODS 5 1. Culture of immortalized human neural stem cells (hNSCs) 5 2. Pretreatment of scaffold and cell seeding 5 3. ELISA of NT-3 6 4. Spinal cord injury and implantation of scaffold 7 5. Histological preparation . 8 6. Immuohistochemistory 8 7. Quantitative morphometry9 III. RESULTS12 1. Seeding NT-3 overexpressing hNSCs in PLGA scaffold 12 2. Implantation of PLGA seeded with hNSCs (PLGA-NT3.F3) 15 3. Detection of human neural stem cell in scaffold 18 4.Deposition of chondroitin sulfate proteoglycans (CSPG) in the lesion 22 5. Remodeling of serotonergic axons 24 IV. DISCUSSION 29 V. CONCLUSION 33 REFERENCES 34 국문요약 43 |LIST OF FIGURES Fig. 1. NT-3 overexpressing human neural stem cells 13 Fig. 2. Seeding human neural stem cells (hNSCs) in PLGA Scaffold 14 Fig. 3. Implantation of PLGA seeded with hNSCs (PLGA-NT3.F3) 16 Fig. 4. Area quantifications of spared tissue and residual scaffold 17 Fig. 5. Detection of transplanted human neural stem cells (hNSCs) 19 Fig. 6. Detection of transplanted human neural stem cells (hNSCs) in white matter 20 Fig. 7. Differentiation of neural stem cells in white matter 21 Fig. 8. Deposition of chondroitin sulfate proteoglycans (CSPGs) at the interface 23 Fig. 9. Detection of serotonergic neuron in lesion site by 5-HT staining 26 Fig. 10. Quantification of serotonergic neurons in caudal area : 5-HT staining 27 |LIST OF ABBREVIATION APC-CC1, anti-APC (Ab-7) mouse mAb (CC-1) BDNF, brain-derived neurotrophic factor CNS, central nervous system CS-56, anti-chondroitin sulfate CSPG, chondroitin sulfate proteoglycans DAPI, 4’, 6 –Diamidino-2-Phenylindole Dihydrochloride DMEM, dulbecco’s modified eagle’s medium DW, distilledwater ELISA, enzyme-linked immunosorbent assay EM, electron microscope F3, human brain F3 GFAP, anti-glial fibrillary acidic protein hNSC, human neural stem cell 5-HT, 5-hydroxytryptamine Hu-Mito, anti-human mitochondria L, left NGF, nerve growth factor NSAID, nonsteroidal antiinflammatory drug NT3, neurotrophin-3 NT3.F3, neurotrophin-3 overexpressing F3 MAP2, anti-microtubule-associated protein 2 PBS, phosphate buffered saline PLA, poly lactic acid PLGA, poly (lactic-co-glycolic acid) PGA, poly (glycolic acid) P/S, penicillin streptomycin R, right ROI, regions of interest RT, room temperature S or SC, scaffold SCI, spinal cord injury T, tissue VH, ventral horn WM, white matter | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 척수재생을 위한 신경줄기세포와 고분자 지지체를 이용한 복합적인 치료전략의 개발 | - |
| dc.title.alternative | Young Mi Kang | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Young Mi Kang | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2009. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 567834 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000009802 | - |
| dc.subject.keyword | spinal cord injury | - |
| dc.subject.keyword | neural stem cell | - |
| dc.subject.keyword | neurotrophin-3 | - |
| dc.subject.keyword | poly (lactic-co-glycolic acid) scaffolds | - |
| dc.subject.keyword | serotonergic axon | - |
| dc.description.alternativeAbstract | ABSTRACT- Development of Combinatorial Strategies Employing Neural Stem Cells and Polymer Scaffold for Spinal Cord Repair Stem cell-based therapy holds promise to enhance functional recovery following spinal cord injury (SCI). Most evidence that transplantation of neural stem cells can produce beneficial outcomes after SCI has been derived from rodent models. Before being translated to human patients, it would be needed to examine the therapeutic effects in larger animal models where weight-bearing locomotion should be much more challenging. The present study examined therapeutic effects of a combinatorial strategy centering on human neural stem cells (hNSCs) in a canine hemisection SCI model. After traumatic injuries to the spinal cord, secondary injuries aggravate the extent of damage, expanding cystic cavities at the lesion site. Cavity wall is surrounded by glial scar and severely impedes axonal regrowth. Therefore, any regenerative strategy could be complemented with an attempt to bridge lesion cavities. In the current study, we implanted poly (lactic-co-glycolic acid) (PLGA) scaffolds to facilitate the delivery of neural stem cells. We sought to enhance therapeutic efficacy by ectopic expression of neurotrophin-3 (NT-3) in the hNSCs. NT-3 overexpressing hNSCs (NT3.F3) were produced by retroviral transduction of the immortalized hNSC line (F3) with human NT-3 cDNA. NT3.F3 cells were seeded into a predesigned PLGA (65:35) scaffolds, and the PLGA scaffolds with hNSCs (PLGA-NT3.F3) were implanted immediately after left hemisection at T11 in female dogs weighing 20-30 kg. The PLGA scaffold seemed to nicely fill the lesion cavity, showing a varying degree of biodegradation by 12 weeks. Survival of grafted cells was confirmed at 2 weeks, and some of the grafted cells migrated to the host tissue. There were very few regenerating axons into the scaffold in both groups. Moreover, the ventral horns caudal to the hemisected region were more profusely innervated by serotonergic axons in animals with PLGA-NT3.F3. These findings raise a possibility that implantation of PLGA-NT3.F3 can produce beneficial motor outcomes by promoting axonal remodeling below the lesion. This study suggests that the therapeutic strategy combining multidisciplinary approaches can be feasible and effective for spinal cord repair in larger species. Key words: spinal cord injury; neural stem cells; NT3; PLGA scaffold; serotonergic axon | - |
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