Cell Signaling Analysis by Mass Spectrometry under Coculture Conditions on an Integrated Microfluidic Device

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• 作者：Huibin Wei ; Haifang Li ; Sifeng Mao ; Jin-Ming Lin
• 刊名：Analytical Chemistry
• 出版年：2011
• 出版时间：December 15, 2011
• 年：2011
• 卷：83
• 期：24
• 页码：9306-9313
• 全文大小：982K
• 年卷期：v.83,no.24(December 15, 2011)
• ISSN：1520-6882

文摘

A microfluidic device was integrated in a controlled coculture system, in which the secreted proteins were qualitatively and semiquantitatively determined by a directly coupled mass spectrometer. PC12 cells and GH3 cells were cocultured under various conditions as a model of the regulation of the organism by the nervous system. A micro-solid phase extraction (SPE) column was integrated in order to remove salts from the cells secretion prior to mass spectrometry detection. A three layer polydimethylsiloxane (PDMS) microfluidic device was fabricated to integrate valves for avoiding contamination between the cells coculture zone and the pretreatment zone. Electrospray ionization (ESI)-quadrupole (Q)-time of flight (TOF)-mass spectrometry was employed to realize highly sensitive qualitative analysis and to implement semiquantitative analysis. Furthermore, cell migrations under various coculture conditions were observed and discussed. The inhibition on growth hormone secretion from GH3 cells by dopamine released from PC12 cells was investigated and demonstrated. Thus, the developed platform provides a useful tool on cell to cell signaling studies for disease monitoring and drug delivery control.