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TNF-α와 IL-1β로 자극 된 태아 연골 유래 줄기 세포에 의한 활막 세포 염증 마커의 하향 조절
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 민병현 | - |
| dc.contributor.author | 신동일 | - |
| dc.date.accessioned | 2019-04-01T16:40:46Z | - |
| dc.date.available | 2019-04-01T16:40:46Z | - |
| dc.date.issued | 2019-02 | - |
| dc.identifier.other | 28844 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/14943 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :분자과학기술학과,2019. 2 | - |
| dc.description.tableofcontents | Contents Abstract--------------------------------------------------------------------------------------------ⅰ Contents------------------ -------------------------------------------------------------------------ⅲ List of Figures------------------------------------------------------------------------------------ⅳ List of Table---------------------------------------------------------------------------------------ⅴ 1. Introduction--------------------------------------------------------------------------------------1 2. Materials and Methods-------------------------------------------------------------------------5 2.1 Isolation and Culture of Sem Cells-----------------------------------------------------5 2.2 Optimization of Priming Condition and Conditioned Media Preparation---------5 2.3 Anti-Inflammation Assay----------------------------------------------------------------6 2.4 RNA Analysis-----------------------------------------------------------------------------6 2.5 PGE2 Quantification----------------------------------------------------------------------6 2.6 Western Blot Analysis--------------------------------------------------------------------7 2.7 Human Cytokine Array------------------------------------------------------------------7 2.8 Cell Morphology Analysis---------------------------------------------------------------8 2.9 Flow Cytometry Analysis----------------------------------------------------------------8 2.10 Multilineage Differentiation-------------------------------------------------------------8 2.11 Statistical Analysis------------------------------------------------------------------------9 3. Results------------------------------------------------------------------------------------------11 3.1 Optimization of priming condition----------------------------------------------------11 3.2 FCSCs primed with TNF-α (10 ng/mL) and IL-1β (15 ng/mL) can down-regulate the level of pro-inflammatory markers in inflammatory synoviocytes----------15 3.3 Determination of protein mediating the anti-inflammatory effects of FCSCs--18 3.4 The stem cell character of FCSCs is maintained after priming-------------------21 4. Discussion-------------------------------------------------------------------------------------25 References------------------------------------------------------------------------------------------28 국문요약-------------------------------------------------------------------------------------------35 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | TNF-α와 IL-1β로 자극 된 태아 연골 유래 줄기 세포에 의한 활막 세포 염증 마커의 하향 조절 | - |
| dc.title.alternative | Dong-Il Shin | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Dong-Il Shin | - |
| dc.contributor.department | 일반대학원 분자과학기술학과 | - |
| dc.date.awarded | 2019. 2 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 905322 | - |
| dc.identifier.uci | I804:41038-000000028844 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/common/orgView/000000028844 | - |
| dc.subject.keyword | stem cell | - |
| dc.subject.keyword | priming | - |
| dc.subject.keyword | anti-inflammatory effect | - |
| dc.description.alternativeAbstract | Abstract PURPOSE. Fetal cartilage-derived stem cells (FCSCs) have been shown to contribute to OA therapy as novel cells with anti-inflammatory properties. However, efforts to enhance the anti-inflammatory ability of FCSCs have not yet been conducted. In this study, the anti-inflammatory properties of FCSCs were increased by priming and evaluated in inflammatory synoviocytes. METHODS. FCSCs were isolated from the fetal cartilage tissue. Priming was carried out by stimulating FCSCs using a priming factor such as Poly(I:C), IFN-γ, TNF-α and IL-1β and cell culture supernatants were obtained as conditioned media (CM). The anti-inflammatory effects of FCSCs were evaluated by treating CM in inflammatory synoviocytes. The evaluation was carried out by confirming the expression of the pro-inflammatory gene TNF-α, IL-1β and IL-6 through qRT-PCR. In addition, the concentration of PGE2 secreted by inflammatory synoviocytes was measured through ELISA. To identify the factors that affect the anti-inflammatory effects of FCSCs, the growth factors (IGF-1, TGF-β and FGF-2) were measured with qRT-PCR and the expression of IDO-1 was confirmed with western blotting. We also identified various secreted factors in the CM of FCSCs through Protein Array. Finally, we analyzed the characteristics of FCSCs using a Microscope, FACS and Histologic analysis to confirm that the stem cell characters are maintained. RESULTS. When TNF-α (10 ng/mL) and IL-1β (15 ng/mL) were treated with FCSCs for 24 h, the anti-inflammatory properties of FCSCs were most significantly increased. In inflammatory synoviocytes treated with CM of primed-FCSCs, the expression of the pro-inflammatory gene TNF-α, IL-1β and IL-6 decreased. In addition, the concentration of PGE2 was significantly reduced in CM of primed-FCSCs. The anti-inflammatory effects of FCSCs were maximized when the concentration of CM was doubled. The expression of IGF-1, TGF-β, FGF-2 and IDO-1 significantly increased after priming, and the Protein Array confirmed an increase in the secretion of more than 20 proteins at the primed-FCSCs. The cell morphology and size did not differ in normal- and primed-FCSCs, and stem cell marker was maintained. Histologic analysis confirmed that the multilineage differentiation potential of FCSCs was maintained. CONCLUSIONS. In conclusion, this study demonstrated that CM of Primed-FCSCs showed higher ability to inhibit inflammatory events in synoviocytes than those of CM of unprimed-FCSCs. Moreover, probably upregulated anti-inflammatory factors mediated anti-inflammatory effects of FCSCs. | - |
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