| Summary: | 碩士 === 國立陽明大學 === 生醫光電研究所 === 101 === This study demonstrates the possibility of using selective plane illumination microscopy (SPIM) to observe live tumor spheroid inside a microfluidic device . SPIM provides optical sectioning capability, low photo toxicity, and low photo-bleaching effect which are useful for studying large specimen like 3-D cell spheroids.
Culture of cells as three-dimensional (3-D) aggregates, such as cancer spheroids, can improve in vitro tests for basic biomedical research and drug testing. In comparison, conventional two dimensional monolayer cultures do not provide the geometrical, biochemical and mechanical factors created in real tissues. We used a simple microfluidic device for the formation and co-culture spheroids of liver carcinoma cells ( HepG2) and human umbilical vein endothelial cells (HUVEC) using gravity driven cell aggregation, and used selective plane illumination microscopy (SPIM) to study the growth of a single spheroid inside the microfluidic device. We could culture the 3-D spheroids inside the device for more than 4 days while exchanging the culture medium. We could measure cell spheroid volume quantitatively and we found out spheroid grow up day by day. We observed that HUVEC inside HepG2 tumor spheroid form approximate circular structure similar to pre tube formation.
Combining microfluidics and SPIM we could study the cell-cell interactions in 3-D spheroids for a long period of time. This study in co-culture spheroids may pave a way to develop new models for in vitro drug testing and tumor angiogenesis.
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