허혈성뇌졸중 쥐에서 신경세포로 분화유도된 중간엽줄기세포의 치료 효과
Therapeutic effect of neural induced mesenchymal stem cells in ischemic rat brain
- 주제(키워드) mesenchymal stem cells , neurogenin-1 , ischemic stroke , Intra-arterial transplantation
- 발행기관 아주대학교
- 지도교수 서해영
- 발행년도 2017
- 학위수여년월 2017. 2
- 학위명 박사
- 학과 및 전공 일반대학원 의생명과학과
- 실제URI http://www.dcollection.net/handler/ajou/000000024882
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Mesenchymal stem cells (MSCs) have been shown to improve a variety of neurological dysfunction by their paracrine effects. Neurogenin-1 (Ngn1) is a proneural gene that directs neuronal differentiation of progenitor cells during development. In recently study, intracranial injection of Ngn1-expessing MSCs showed the remarked improvement of motor dysfunction in stroke model compared to MSC and PBS treated group. However, intracranial injection is not feasible method to use in clinical field. Therefore, we conducted the study to investigate that intra-arterial injection of Ngn1-expressing MSCs can improve motor deficit in ischemic rat model. Cerebral ischemic stroke is a serious public health concern. It causes considerable death and disability. Only limited treatment options are available in the acute phase of stroke. Fifteen Sprague-Dawley rats were subinjected to transient middle cerebral artery occlusion (tMCAo) of 2 hours with the suture occlusion model. Magnetic resonance image (MRI), including diffusion-weighted imaging (DWI) and T2-weighted imaging was performed at 2, 7 and 28days after withdrawal of the suture. Motor function evaluation including ratarod test and adhesive removal test was performed at 1, 7, 14 and 28 days. Amimals were divided into 3 subgroups. Each group received 1×10⁶ MSC-Ngn1 cells, 1×10⁶ MSC-lacZ cells and normal saline respectively. The distribution and phenotype of injected stem cells were compared among the groups. Rat injected with MSC-Ngn1 showed the tendency of motor dysfunction improvement compared to MSC-LacZ and control groups. The induction of neural stem cell number was greater in MSC-Ngn1 injected rat than in other groups. Intra-arterial injection of MSC-Ngn1 cell in stroke model showed the remarkable improvement of motor dysfunction. Intra-arterial injection can be the feasible method of stem cell transplant.
more목차
Part. I. Comparison of MSC-Neurogenin1 administration modality in MCAo rat model 1
I. INTRODUCTION 2
II. MATERIALS AND METHODS 4
A. MSCs/Ngn1 preparation 4
B. Induction of ischemic models 5
C. Transplantation 8
D. Behavioral testing 10
E. Infarct volume measurement 11
F. In vivo tracking of superparamagnetic iron oxide (SPIO)-labeled cells 12
G. Histological analysis 13
H. Statistical analysis 14
III. RESULTS 15
A. Therapeutic effects of MSCs/Ngn1 according to administration modality 16
B. Serial changes in infarct volume according to administration modality 17
C. MSCs/Ngn1 distribution according to administration modality 19
D. MSCs/Ngn1 transdifferentiation in a rat stroke model 21
IV. DISCUSSION 23
Part. II. Therapeutic Effects of Intra-Arterial Delivery of neural-induced mesenchymal stem cells in The Ischemic Brain 27
I. INTRODUCTION 28
II. MATERIALS AND METHODS 29
A. Cell preparation and culture 29
B. Transient middle cerebral artery occlusion (MCAo) model 29
C. Cell administration 30
D. Measurement of infarct volume 32
E. SPIO labeling of cells 33
F. In Vivo MRI cell tracing 34
G. Cell Migration Assays 35
H. RNA isolation and RT-qPCR 36
I. Immunohistochemistry 38
J. Statistical Analysis 39
III. RESULTS 40
A. IA transplantation of MSC-ngn1 improve functional recovery 40
B. MRI Tracking of cells after IA TP 43
C. Cell migration mechanism 49
D. Paracrine effect of MSC-ngn1 cells on ischemic brain 52
E. Tissue integrity was preserved in the MSC-Ngn1 treated brain 56
F. Transdifferentiation of MSC-Ngn1 Cells In Vivo 58
IV. DISCUSSION 60
V. CONCLUSION 63
VI. REFERENCE 64
국문요약 69
