請用此 Handle URI 來引用此文件:
http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17723
標題: | 間葉幹細胞與癌細胞或纖維母細胞在幾丁聚醣上3D共培養系統的CD44表現 CD44 expression of MSC and Cancer Cell or Fibroblast in 3D Co-culture System on Chitosan Film |
作者: | Ching-Wen Tsai 蔡靜雯 |
指導教授: | 楊台鴻(Tai-Horng Young) |
關鍵字: | 幾丁聚醣,共培養,CD44,間葉幹細胞,癌細胞,纖維母細胞, chitosan,co-culture,CD44,mesenchymal stem cells,cancer cells,fibroblasts, |
出版年 : | 2013 |
學位: | 碩士 |
摘要: | 本研究使用間葉幹細胞3A6、癌細胞SW620和纖維母細胞Hs68進行實驗。首先,利用流式細胞儀和分化測試驗證細胞特性,證明間葉幹細胞3A6具有類似間葉幹細胞的特性,癌細胞可用抗原標記標定出癌幹細胞。
幾丁聚醣是一個天然、具生物可分解、生物相容性、無生物毒性,且經由美國食品藥品管理局認可的多醣類,本研究將其塗佈於基材上培養細胞。當間葉幹細胞、癌細胞和纖維母細胞分別種植於幾丁聚醣基材上時,細胞都會呈現懸浮且聚集的型態。利用穿膜醣蛋白CD44抗原標記,比較細胞在不同培養材料上的表現,和推測其可能影響原因。 接著,將此三種細胞在幾丁聚醣基材上利用直接共培養系統培養,探討細胞之間的交互作用。把細胞分別用螢光標定後,以倒立式螢光顯微鏡觀察細胞在基材上共培養後的形態;更進一步,利用共軛焦螢光顯微鏡證實3D細胞球的排列組合。並利用細胞膜上的穿膜醣蛋白CD44檢視共培養系統和高分子材料幾丁聚醣對CD44抗原標記的影響。 最後,利用MTT assay、免疫螢光染色Ki-67和LIVE/DEADR Viability/Cytotoxicity Assay Kit,相互佐證和探討細胞在細胞球上不同位置的狀態。 In this study, we used mesenchymal stem cells (MSCs) 3A6, cancer cells SW620 and fibroblasts Hs68. First, we demonstrated that 3A6 is similar with MSCs by flow cytomerty analysis and differentiation assay. In addition, antigen markers were used to mark cancer stem cells. Chitosan is a natural, biodegradable, biocompatible, non-toxic and U.S. Food and Drug Administration (FDA) approved polysaccharide. In this study, chitosan was used as the coating substrates. When mesenchymal stem cells, cancer cells and fibroblasts were cultured on chitosan substrates, all cells became suspended and aggregated into spheroids. We used the antigens of transmembrane glycoprotein CD44 on the cell membrane to mark cells in order to compare the influence of different materials, and determine the causes of any discrepancies. Furthermore, we cultured MSCs, cancer cells, and fibroblasts on chitosan substrates in direct co-culture systems to explore the interactions between the three types of cells. We labeled these cells by the CellTrackerTM and used the inverted fluorescence microscope to observe the morphology and distribution of cells on substrates. In addition, we used confocal fluorescence microscope to check the arrangement of 3D spheroids. Besides, we used the antigens of transmembrane glycoprotein CD44 to mark cells in order to observe the effect of different substrates on the co-culture systems. Finally, we used the MTT assay, immunofluorescence staining Ki-67 and LIVE/DEADR Viability/Cytotoxicity Assay Kit to prove and discuss the status of cells on different positions of cell spheroids. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/17723 |
全文授權: | 未授權 |
顯示於系所單位: | 醫學工程學研究所 |
文件中的檔案:
檔案 | 大小 | 格式 | |
---|---|---|---|
ntu-102-1.pdf 目前未授權公開取用 | 5.94 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。