An In-vitro Functional Assay to Assess Migration of Cancer Cells using the Micro Gap Plate

• Main Authors: Chia-Hao Yao, 游嘉豪
• Other Authors: 胡文聰
• Format: Others
• Language: en_US
• Published: 2016
• Online Access: http://ndltd.ncl.edu.tw/handle/03128542308454676535

碩士 === 國立臺灣大學 === 應用力學研究所 === 104 === In-vitro chemotaxis studies of tumor cells facilitate understanding of tumor progression and assessment of aggressiveness. This work presents the use of a SBS-compatible 96-well plate with special microfluidic gap feature in each well to characterize the aggressiveness of tumor cells with a custom-tailored chemotaxis gradient for each well. This microgap plate (MGP) is also amenable towards imaging under microscopy. The MGP was characterized with three breast cancer cell lines (MCF7, MDA-MB-231 and HS578T) and two types of patient-derived culture cells, or PDCC (ABC72PE and ABC37T). The cell lines are selected based on their phenotypes: epithelial (MCF7), intermediate epithelial-mesenchymal (MDA-MB-231), and mesenchymal (HS578T). The PDCC were cultured from biopsies from metastatic sites of breast cancer patients, with epithelial (ABC72PE) and intermediate epithelial-mesenchymal (ABC37T) phenotypes. Microenvironment with chemotaxis gradients of epidermal growth factor (EGF) and hepatocyte growth factor (HGF) (0, 25, 50 and 100 ng/ml) were tested. Results confirm migration ability of all cell lines exhibit directional movement toward higher EGF/HGF concentration. Further, migration ability of MDA-MB-231 cells with intermediate epithelial-mesenchymal phenotype is more aggressive than either the standalone epithelial (MCF7) or mesenchymal (HS578T) phenotype. Similarly, this intermediate phenotypical characteristic also holds for patient-derived cells where ABC37T exhibits more aggressive behavior than ABC72PE cells. The 96-well high-throughput MGP is shown to be a versatile cellular assay to assess tumor aggressiveness under custom-tailored chemotaxis gradient in each well and might be amenable towards functional assessment of clinical tissues.