김병곤 김미경 2007-08 2672 https://aurora.ajou.ac.kr/handle/2018.oak/17166 학위논문(석사)----아주대학교 일반대학원 일반대학원 :의학과,2007. 8 국문초록 Ⅰ. INTRODUCTION = 1 Ⅱ. MATERIALS AND METHODS = 5 1. Cell culture = 5 2. Generation of human NSC F3.Akt1 cell line = 5 3. RT-PCR analysis = 6 4. Treatment of H₂O₂ = 6 5. Cell viability assay = 7 6. Western blot analysis = 7 7. Oxygen glucose deprivation (OGD) experiment = 8 8. Mouse Intracerebral Hemorrhage (ICH) model = 8 9. Cell transplantation = 9 10. Behavioral Testing = 9 11. Histology and immunohistochemistry = 10 12. Stereological cell counts = 11 13. Statistical analysis = 12 Ⅲ. Results = 13 1. Stable human neural stem cell line over-expressing Akt1 = 13 2. F3.Akt1 cells prevent H2O2-Induced cell death = 16 3. Oxygen glucose deprivation (OGD) experiment = 18 4. Functional recovery in ICH animals by hNSC transplantation = 20 5. Transplanted hNSCs differentiate into neurons and astrocytes = 22 6. Survival of transplanted F3 and F3.Akt1 hNSCs in ICH brain = 25 Ⅳ. DISCUSSION = 27 Ⅴ. CONCLUSION = 32 REFERENCES = 33 국문 요약 = 43 eng The Graduate School, Ajou University 아주대학교 논문은 저작권에 의해 보호받습니다. Akt1 과발현을 유도한 인간 신경줄기 세포주의 마우스 뇌출혈 뇌졸중 모델에서의 치료 효과 Kim, Mi-Kyung Thesis 아주대학교 일반대학원 Kim, Mi-Kyung 일반대학원 의학과 2007. 8 Master http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000002672 human neural stem cells transplantation Akt1 마우스 뇌출혈 뇌졸중 인간 신경줄기 Akt1 (protein kinase B), a serine/threonine kinase, has emerged as a critical molecule in signal transduction pathways involved in cell proliferation, growth, survival, apoptosis and glucose metabolism. A previous study has shown that transplanted human neural stem cells (hNSCs) selectively migrate to the brain and induce behavioral recovery in mouse intracerebral hemorrhage (ICH) stroke model. However, many of the grafted hNSCs did not survive the transplantation, which may have reduced therapeutic potential. In this study, we postulated that hNSCs over-expressing Akt1 transplanted into the lesion could improve survival of grafted hNSCs and behavioral recovery in mouse ICH model. To establish Akt1 over-expressing hNSC line, F3 hNSCs were transduced with retroviral vectors containing mouse Akt1 cDNA. We examined the viability of Akt1 over-expressing hNSCs against to the H2O2-induced cell death and oxygen glucose deprivation (OGD) condition in vitro. F3.Akt1 cells were showed more resistance to the cell death stimuli. To investigate the therapeutic effects in vivo, ICH was induced in adult mice by unilateral injection of bacterial collagenase into striatum and then, we transplanted F3 and F3.Akt1 hNSCs into the animal brain. Transplantation of F3.Akt1 hNSCs increased survival of grafted cells by 0.5 ？ 2 fold at two weeks and eight weeks, and induced behavioral improvement in animals. These results demonstrate that the over expression of Akt1 supports the prolonged survival of the engrafted cells and protects hNSCs from cell death by the inhibitory death machinery activation.