title: A PDMS-Based Microfluidic Hanging Drop Chip for Embryoid Body Formation.
abstract: The conventional hanging drop technique is the most widely used method for embryoid body (EB) formation. However, this method is labor intensive and limited by the difficulty in exchanging the medium. Here, we report a microfluidic chip-based approach for high-throughput formation of EBs. The device consists of microfluidic channels with 6 × 12 opening wells in PDMS supported by a glass substrate. The PDMS channels were fabricated by replicating polydimethyl-siloxane (PDMS) from SU-8 mold. The droplet formation in the chip was tested with different hydrostatic pressures to obtain optimal operation pressures for the wells with 1000 μm diameter openings. The droplets formed at the opening wells were used to culture mouse embryonic stem cells which could subsequently developed into EBs in the hanging droplets. This device also allows for medium exchange of the hanging droplets making it possible to perform immunochemistry staining and characterize EBs on chip.
authors.affiliation: Institutes of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan 35053, Taiwan. fecancer@gmail.com.
authors.forename: Huei-Wen
authors.initials: HW
authors.lastname: Wu
authors.affiliation: Institute of Nano Engineering and MicroSystems, National Tsing Hua University, Hsinchu 30013, Taiwan. top.study@kimo.com.
authors.forename: Yi-Hsing
authors.initials: YH
authors.lastname: Hsiao
authors.affiliation: Institute of Nano Engineering and MicroSystems, National Tsing Hua University, Hsinchu 30013, Taiwan. chihchen@mx.nthu.edu.tw.|Department of Power Mechanical Engineering, National Tsing Hua University, Hsinchu 30013, Taiwan. chihchen@mx.nthu.edu.tw.
authors.forename: Chih-Chen
authors.initials: CC
authors.lastname: Chen
authors.affiliation: Institute of Cellular and System Medicine, National Health Research Institutes, Zhunan 35053, Taiwan. syet@nhri.org.tw.
authors.forename: Shaw-Fang
authors.initials: SF
authors.lastname: Yet
authors.affiliation: Institutes of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Zhunan 35053, Taiwan. chsu@nhri.org.tw.|Institute of Nano Engineering and MicroSystems, National Tsing Hua University, Hsinchu 30013, Taiwan. chsu@nhri.org.tw.
authors.forename: Chia-Hsien
authors.initials: CH
authors.lastname: Hsu
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002460:Cell Line; D058732:Embryoid Bodies / Q000166:cytology*; D053595:Embryonic Stem Cells / Q000166:cytology*; D004867:Equipment Design; D007150:Immunohistochemistry; D051379:Mice; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*
keywords: embryoid body; embryonic stem cell; microfluidic hanging drop