title: Integrated microdevice for long-term automated perfusion culture without shear stress and real-time electrochemical monitoring of cells.
abstract: Electrochemical techniques based on ultramicroelectrodes (UMEs) play a significant role in real-time monitoring of chemical messengers' release from single cells. Conversely, precise monitoring of cells in vitro strongly depends on the adequate construction of cellular physiological microenvironment. In this paper, we developed a multilayer microdevice which integrated high aspect ratio poly(dimethylsiloxane) (PDMS) microfluidic device for long-term automated perfusion culture of cells without shear stress and an independently addressable microelectrodes array (IAMEA) for electrochemical monitoring of the cultured cells in real time. Novel design using high aspect ratio between circular "moat" and ring-shaped micropillar array surrounding cell culture chamber combined with automated "circular-centre" and "bottom-up" perfusion model successfully provided continuous fresh medium and a stable and uniform microenvironment for cells. Two weeks automated culture of human umbilical endothelial cell line (ECV304) and neuronal differentiation of rat pheochromocytoma (PC12) cells have been realized using this device. Furthermore, the quantal release of dopamine from individual PC12 cells during their culture or propagation process was amperometrically monitored in real time. The multifunctional microdevice developed in this paper integrated cellular microenvironment construction and real-time monitoring of cells during their physiological process, and would possibly provide a versatile platform for cell-based biomedical analysis.
authors.affiliation: Key Laboratory of Analytical Chemistry for Biology and Medicine (Ministry of Education), College of Chemistry and Molecular Sciences, Wuhan University, Wuhan, China.
authors.forename: Lin-Mei
authors.initials: LM
authors.lastname: Li
authors.forename: Wei
authors.initials: W
authors.lastname: Wang
authors.forename: Shu-Hui
authors.initials: SH
authors.lastname: Zhang
authors.forename: Shi-Jing
authors.initials: SJ
authors.lastname: Chen
authors.forename: Shi-Shang
authors.initials: SS
authors.lastname: Guo
authors.forename: Olivier
authors.initials: O
authors.lastname: Français
authors.forename: Jie-Ke
authors.initials: JK
authors.lastname: Cheng
authors.forename: Wei-Hua
authors.initials: WH
authors.lastname: Huang
mesh_terms: D000818:Animals; D001331:Automation; D018929:Cell Culture Techniques; D002460:Cell Line; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004298:Dopamine / Q000032:analysis; D055664:Electrochemical Techniques; D006801:Humans; D008839:Microelectrodes; D046210:Microfluidic Analytical Techniques; D010477:Perfusion; D051381:Rats; D033081:Shear Strength
chemical_list: D004129:Dimethylpolysiloxanes; C013830:baysilon; D004298:Dopamine