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급성 brain trauma 모델에서 111In으로 표지된 중간엽 줄기세포의 귀소성
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 윤준기, 안영환 | - |
| dc.contributor.author | Park, Bok-Nam | - |
| dc.date.accessioned | 2019-10-21T07:14:27Z | - |
| dc.date.available | 2019-10-21T07:14:27Z | - |
| dc.date.issued | 2010-08 | - |
| dc.identifier.other | 10981 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/17663 | - |
| dc.description | 학위논문(박사)--아주대학교 일반대학원 :의학과,2010. 8 | - |
| dc.description.tableofcontents | ABSTRACT ----------------------------------------- i TABLE OF CONTENTS ------------------------------ iv LIST OF FIGURES ----------------------------------- vi I. INTRODUCTION ---------------------------------- 1 II. MATERALS AND METHODS ----------------------- 3 A. Isolation and culture of rat BMSCs ----------------- 3 B. Generation of an animal model of acute brain trauma 4 C. Synthesis and radioabeling of mesenchymal stem cells with 111In-tropolone ------------------------------- 4 D. In vitro stability and cell viability of mesenchymal stem cells with 111In-tropolone-------------------------- 5 E. Dose-dependent effect of 111In on the growth of BMSCs ---------------------------------------- 6 F. In vivo tacking of 111In-BMSCs by gamma camera in animal models ----------------------------------- 6 G. PKH 26 labeling of mesenchymal stem cells -------- 7 H. Tissue preparation with DAPI staining for confocal microscopy -------------------------------------- 7 I. In vitro BrdU labeling for 111In-BMSCs -------------- 8 J. Annexin V-FITC/PI double staining for 111In-BMSCs 9 K. Cytochemical staining with SA-β-galactosidase for 111In-BMSCs ---------------------------------- 9 L. Statistical analysis -------------------------------- 10 III. RESULTS --------------------------------------- 11 A. Radiolabeling efficiency and viability of 111In-BMSCs 11 B. Dose-dependent growth of 111In-BMSCs ----------- 12 C. In vivo tracking of 111In-BMSCs by gamma camera in trauma models and controls --------------------- 13 D. Histological analysis of transplanted BMSCs in animal model of trauma ---------------------------------- 14 E. Cell cycle analysis by flow cytometry -------------- 16 F. Annexin V-FITC/PI double staining flow cytometry -- 18 G. Senescence-associated-β-galactosidase histochemistry ------------------------------------ 20 IV. DISCUSSION ------------------------------------ 22 V. CONCLUSION ----------------------------------- 29 REFERENCES -------------------------------------- 30 국문요약 -------------------------------------------- 38 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | 급성 brain trauma 모델에서 111In으로 표지된 중간엽 줄기세포의 귀소성 | - |
| dc.title.alternative | Homing of 111In-Labeled Bone Marrow Mesenchymal Stem Cells in Acute Brain Trauma Model | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.department | 일반대학원 의학과 | - |
| dc.date.awarded | 2010. 8 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 568772 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/jsp/common/DcLoOrgPer.jsp?sItemId=000000010981 | - |
| dc.subject.keyword | In-111 tropolone | - |
| dc.subject.keyword | Bone marrow mesenchymal stem cells | - |
| dc.subject.keyword | Cell tracking | - |
| dc.subject.keyword | Growth arrest | - |
| dc.description.alternativeAbstract | This study was to evaluate the in vivo distribution of intravenously transplanted bone marrow-derived mesenchymal stem cells (BMSCs) in an acute brain trauma model by 111In-tropolone labeling and to perform the effect of 111In-labeling on the viability and functions of BMSCs. Rat BMSCs were labeled with 37 MBq 111In-tropolone. Their labeling efficiency and in vitro retention rate were measured. To evaluate dose-dependent effect of 111In-labeling, BMSCs were labeled with various doses (0.4-11.1 Bq/cell) of 111In-tropolone, and growth curve analysis, fluorescent activated cell sorter (FACS) analysis after staining with 5-bromo-2-deoxy-uridine (BrdU), and microscopic evaluation after 5-bromo-4-chloro-3-indolyl-D-galactopyranoside (X-gal) staining were performed until the 14th day. FACS analysis after staining with Annexin V- fluorescein isothiocyanate (FITC) and propidium iodide (PI) was performed at early (3 and 12 hr) and late (7 days) stages with higher doses of 111In (11.1 and 33.3 Bq/cell) to evaluate apoptotic or necrotic change of labeled BMSCs. The biodistribution of 111In-BMSCs in trauma models was compared with those in sham-operated rats and normal rats by gamma camera images. The migration of 111In-BMSCs to the traumatic brain was evaluated using confocal microscope. The labeling efficiency of 111In-BMSCs was 66 ± 5%, and their retention rate was 85.3% at 1 h after labeling. There was no difference in the number of viable cells between 111In-BMSCs and controls at 48 h after labeling. However, the proliferation of 111In-BMSCs was inhibited after the third day of labeling, and it did not reach confluency. For lower doses of 111In (0.4 and 1.1 Bq/cell), the growth of labeled stem cells was not significantly different from that of control, whereas, labeling with higher doses of 111In (4.4 and 11.1 Bq/cell) led to a significant proliferative inhibition from the 3rd day to the 14th day. FACS analysis also revealed less BrdU positive cells in BMSCs labeled with 1.1, 4.4 and 11.1 Bq/cell compared with controls on the 3rd day after labeling. Of these, the patterns of cell cycle in BMSCs labeled with 0.4 and 1.1 Bq/cell of 111In were restored similar to controls on the 14th day. On the contrary, BMSCs labeled with 4.4 and 11.1 Bq/cell of 111In could not recover from cell cycle arrest. Senescence-associated β-gal (SA- β-gal) staining was not prominent in all concentrations until the 14th day after labeling. FACS analysis with Annexin V-FITC and PI also revealed no significant apoptosis or necrosis in both early and late stages. On gamma camera images, most of the 111In-BMSCs uptake was observed in the liver and spleen at the second day of injection. The brain uptake of 111In-BMSCs was more prominent in trauma models (1.4%) than in sham-operated (0.5%) or normal rats (0.3%). Radiolabeled BMSCs were observed at the marginal region of traumatic brain on the confocal microscope. We observed the dose-dependent growth inhibition of BMSCs by 111In-labeling, which was developed by dose-dependent, transient cell cycle arrest, not by cellular senescence or apoptosis/necrosis. Although growth inhibition by 111In-labeling need to be evaluated further prior to use in humans, 111In-BMSCs are useful for the tracking of intravenously transplanted mesenchymal stem cells in brain disease models. | - |
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