Design of a Modified Microcontact Printing Device for Patterning Neuronal Network

• Main Authors: Ya-Szu Lin, 林亞思
• Other Authors: Jia-Jin Chen
• Format: Others
• Language: en_US
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/08251033986827334194

碩士 === 國立成功大學 === 醫學工程研究所碩博士班 === 94 === It is essential to culture neuronal cells on the geometric patterned substrate to observe the behavior of neuronal cells. Recent studies have made it possible to control the positioning and outgrowth of neuronal cells by using the microcontact printing (μCP) technique. However, currently available devices are either too expensive or not easy to dismantle. The aim of this study is to design an inexpensive microscopy-based μCP device for aligning the pattern to the substrate to guide neuronal growth. First, procedures for fabrication of the master mold and formation of polydimethylsiloxane (PDMS) stamp were developed. The stamp was inked with the solution of poly-D-lysine (PDL), a cell adhesion factor, to define the cytophilic region. Meanwhile, the stamped substrate was immersed to bovine serum albumin (BSA) for defining the cytophobic region. To align the stamp to microelectrodes in the substrate, image processing techniques were applied on-line to adjust the tilting of stamp and alignment of pattern. Moreover, cell lines and cortical neurons were cultured on the patterned substrates from which general guidelines for viable patterned culturing was investigated. These guidelines found include (1) neurons and cells cannot grow well in a small cytophilic region, e.g., in a cytophilic / cytophobic ratio less than 0.0017; (2) the larger the width of pattern line is, the better the cells can grow; (3) varied cell lines and neurons respond differently to cytophobic reagents, e.g., common use of BSA is proved not effective to repel fibroblasts. Our novel design of microscope-based μCP device provides an effective but inexpensive tool to precisely align the patterns. Further works are needed to follow the general guidelines for systematically developing viable μCP techniques for varied cells which would become a precious tool for studying in-vitro patterns of neuronal networks for neuroelectrophysiological studies.