請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17738
標題: | 胎盤絨毛膜蛻膜間葉細胞效能評估之研究 The Potency Evaluation of Human Placenta Choriodecidual-derived Mesenchymal Stromal Cells |
作者: | Tze-Jung Chen 陳姿蓉 |
指導教授: | 林泰元(Thai-Yen Ling) |
關鍵字: | 胎盤絨毛膜蛻膜間葉細胞,效能評估,功能標記,細胞藥物,免疫調控, Placenta-choriodecidual derived mesenchymal stromal cells,Potency eveluation,Functional marker,Cell drug,Immune modulation, |
出版年 : | 2015 |
學位: | 碩士 |
摘要: | 間葉細胞(MSCs)在過去的研究中被證實能調節生物體內的免疫平衡以適應環境改變,並且能有效地抑制過度的發炎反應。而這些功能使得MSCs在近年來逐漸發展成為細胞藥物,並且被廣泛地運用在臨床治療研究當中。研究指出,細胞藥物會由於來源、捐贈者、培養環境等各種因素,造成治療功能的差異。然而MSCs作為一個細胞藥物,目前並沒有一套標準的評估方式可以判斷MSCs的品質。因此,我們的目標是建立一個標準的in vitro模式以評估MSCs的免疫調節功能,並找到與其相關的功能標記(functional markers)。在我們的研究中利用抗體anti-CD3/28活化小鼠脾細胞的增生,結果發現胎盤絨毛膜蛻膜間葉細胞(placenta-choriodecidual derived mesenchymal stromal cells, pcMSCs) 能有效地抑制脾細胞增生,並且其抑制能力隨著細胞比例增加而上升。我們因此利用pcMSCs對小鼠脾細胞增生的抑制程度,作為評估pcMSCs免疫調控的標準。在評估的過程中,我們發現不同批次的差異以及冷凍保存細胞都會對pcMSCs造成功能影響。其中,來自捐贈者N166和剛解凍的pcMSCs都無法抑制小鼠脾細胞的增殖。由於MSCs可以透過各種細胞激素的分泌及受體表現,來達到免疫調控的能力。我們因此透過RT-PCR評估不同pcMSCs之間免疫調控因子的mRNA表現,研究結果發現功能正常的pcMSCs具有較高的IL-6和COX2表現量。在蛋白質表現部分,ELISA試驗證實psMSCs的IL-6分泌量,與其對脾細胞的抑制能力呈現正相關性。而Western blot試驗則顯示正常功能的pcMSCs具有較高的COX2表現量。此外,我們發現利用胎牛血清(FBS)對功能缺陷的pcMSCs進行培養,能有效地恢復其免疫調控功能。我們的研究證實小鼠脾細胞增生模式能有效地幫助我們評估pcMSCs免疫調控功能的差異,並且IL6和COX2等因子具有作為pcMSCs功能標記的潛力。 Mesenchymal stromal cells (MSCs) are multipotent progenitor cells which are considered a promising tool for cell therapy in recent years. MSCs have been identified to possess immune-modulatory functions on inhibiting pro-inflammatory immune responses and modifying tissue homeostasis to adopt enhancing anti-inflammatory phenotype. According to these properties, they are currently being administrated to patients in clinical trials including graft-versus-host disease (GvHD), bone/cartilage disease or heart disease. Different sources, donors and culture conditions significantly influence the therapeutic potential of MSCs. However, there are no appropriate potency assay to predict the efficacy of MSCs before clinical use. Therefore, we aimed to create a standardized in vitro assay and find potential functional markers to evaluate the potency of pcMSCs. In this study, we stimulated mouse splenocytes proliferation using anti-CD3/28 and co-cultured splenocytes with placenta-choriodecidual derived mesenchymal stromal cells (pcMSCs) which exhibited dose-dependently ability to suppress splenocytes proliferation. We thus measured immune-modulatory efficacy of pcMSCs with this in vitro assay. Results showed that different batch and cryopreservation would affect pcMSCs. Batch N166 and freshly thawed pcMSCs have impaired immunosuppression ability. RT-PCR assays of pcMSCs shown normal pcMSCs have significantly higher level of IL-6 and COX2 mRNA compared to impaired pcMSCs. Protein levels measured by ELISA and western blot showed the same result. Also, we found out that culturing impaired pcMSCs with FBS could reversed the immunosuppressing properties effectively. Our study indicated that the potency of pcMSCs could be evaluated through mouse splenocytes proliferation assay. Importantly, IL6 and COX2 would be potential functional markers of pcMSCs. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17738 |
全文授權: | 未授權 |
顯示於系所單位: | 藥理學科所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-104-1.pdf 目前未授權公開取用 | 2.64 MB | Adobe PDF |
系統中的文件,除了特別指名其著作權條款之外,均受到著作權保護,並且保留所有的權利。