CD44 expression of MSC and Cancer Cell or Fibroblast in 3D Co-culture System on Chitosan Film

• Main Authors: Ching-Wen Tsai, 蔡靜雯
• Other Authors: Tai-Horng Young
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/13299022315318496906

碩士 === 國立臺灣大學 === 醫學工程學研究所 === 101 === 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.