Pneumatic microvalves controlled cell culture chip for the studies on cell-cell interactions

• Main Authors: Yu-Wei Tsao, 曹育維
• Other Authors: Shih-Hao Huang
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/11779902489643093102

碩士 === 國立臺灣海洋大學 === 機械與機電工程學系 === 97 === How cancer cells influence extracellular matrix and interact with nearby normal cells and exchange information through the medium is still a popular research topic at present. By controlling the components in media, adding specific growth factors or stimuli, we can understand the change of cell micro-environments after being influenced, and clarify the mechanisms or signal pathways of which cancer cells interact with normal cells. In this thesis, we develop a microfluidic cell culture chip that contains two cell chambers to co-culture cancer cells and normal fibroblasts. The cell culture chambers are coated with fibronectin on the surface such that the cells are able to attach stably. The fibronectin coating also provides an environment close to those in real tissues. This microfluidic cell culture chip combines with pneumatic microvalves that isolate the chip into two independent culture environments by stopping the flow of the culture medium between the two chambers. In this work, the cell lines are the human lung adenocarcinoma cell CL1-5 with high migration ability, and the human foreskin fibroblast HS68. We control the opening of the microvalves and the flowing directions of the culture medium to study the interactions between these two cells by the secretions of their growth factors and stimuli in the flowing culture medium. Using the cell culture chip, we find that the original culture medium of fibroblasts cannot induce observable change of migration speed of the cancer cells. However, after the fibroblasts are stimulated by the culture medium of cancer cells, the culture medium of fibroblasts can increase the migration speed of the cancer cells. On the other hand, the culture medium of cancer cells directly increases the aspect ratio of fibroblasts. This morphological change is related to the activation of fibroblasts into myofibroblasts. Our design of microfluidic cell culture chip takes the advantages of easy and rapid fabrication. The dimension of the chip fits an on-microscope-stage cell incubator, and hence the experiments can be conducted for a long period. In the future, the chip can be further improved by combining multiple microvalves and complex microfluidic structures as to meet various purposes for cell co-culture experiments.