Cell signaling analysis by mass spectrometry under coculture conditions on an integrated microfluidic device.
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.
Beijing Key Laboratory for Analytical Methods and Instrumentation, Department of Chemistry, Tsinghua University, Beijing, China.
Huibin
H
Wei
Haifang
H
Li
Sifeng
S
Mao
Jin-Ming
JM
Lin
D000818:Animals; D018920:Coculture Techniques; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004298:Dopamine / Q000032:analysis* / Q000302:isolation & purification; D013006:Growth Hormone / Q000032:analysis / Q000302:isolation & purification; D007649:Ketamine / Q000031:analogs & derivatives / Q000494:pharmacology; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D016716:PC12 Cells; D051381:Rats; D015398:Signal Transduction; D052617:Solid Phase Microextraction; D021241:Spectrometry, Mass, Electrospray Ionization
D004129:Dimethylpolysiloxanes; C013830:baysilon; D007649:Ketamine; D013006:Growth Hormone; D004298:Dopamine; C033419:norketamine