title: Microfluidic devices for construction of contractile skeletal muscle microtissues.
abstract: Cell-culture microchips mimicking tissue/organ-specific functions are required as alternatives to animal testing for drug discovery and disease models. Although three-dimensional (3D) cell culture microfluidic devices can create more biologically relevant cellular microenvironments and higher throughput analysis platforms of cell behavior than conventional techniques, devices for skeletal muscle cells have not been developed. In the present study, we aimed to develop microfluidic devices for 3D cultures of skeletal muscle cells. Skeletal muscle cells mixed with a collagen type-I solution was introduced into the microchannel for cells (MC-C) and was gelated. Then, the medium was introduced into the microchannel for medium (MC-M). During this process, connecting microchannels (Con-MCs) prevented leakage of the collagen solution mixed with cells from MC-C to MC-M and supplied the nutrients from the medium in MC-M to the cells in MC-C. Skeletal muscle microtissues cultured in the microchannel for a week consisted of myotubes were confirmed by histological analysis and immunofluorescence staining. The skeletal muscle microtissues in the microchannel contracted in response to externally applied electrical stimulation (1 and 50 Hz). These results indicate that the functional skeletal muscle microtissues were constructed in the microchannel. Thus, the microfluidic device for culturing 3D skeletal muscle microtissues presented in this study has a potential to be used for drug discovery and toxicological tests.
authors.affiliation: Institute for Innovative NanoBio Drug Discovery and Development, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-shimoadachi, Sakyo-ku, Kyoto 606-8501, Japan; Ritusmeikan Global Innovation Research Organization (R-GIRO), Ritsumeikan University, Noji-higashi, Kusatsu, Shiga 525-8577, Japan. Electronic address: shimizu@nubio.nagoya-u.ac.jp.
authors.forename: Kazunori
authors.initials: K
authors.lastname: Shimizu
authors.affiliation: Graduate School of Science and Engineering, Ritsumeikan University, Noji-higashi, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Araki
authors.affiliation: Graduate School of Science and Engineering, Ritsumeikan University, Noji-higashi, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Kohei
authors.initials: K
authors.lastname: Sakata
authors.affiliation: Department of Mechanical Engineering, Ritsumeikan University, Noji-higashi, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Wataru
authors.initials: W
authors.lastname: Tonomura
authors.affiliation: Institute for Innovative NanoBio Drug Discovery and Development, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-shimoadachi, Sakyo-ku, Kyoto 606-8501, Japan; Department of Drug Delivery Research, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-shimoadachi, Sakyo-ku, Kyoto 606-8501, Japan.
authors.forename: Mitsuru
authors.initials: M
authors.lastname: Hashida
authors.affiliation: Institute for Innovative NanoBio Drug Discovery and Development, Graduate School of Pharmaceutical Sciences, Kyoto University, Yoshida-shimoadachi, Sakyo-ku, Kyoto 606-8501, Japan; Ritusmeikan Global Innovation Research Organization (R-GIRO), Ritsumeikan University, Noji-higashi, Kusatsu, Shiga 525-8577, Japan; Department of Mechanical Engineering, Ritsumeikan University, Noji-higashi, Kusatsu, Shiga 525-8577, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Konishi
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation* / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism; D055808:Drug Discovery; D004558:Electric Stimulation; D004867:Equipment Design; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D009119:Muscle Contraction; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism; D018482:Muscle, Skeletal / Q000033:anatomy & histology / Q000166:cytology* / Q000254:growth & development* / Q000378:metabolism; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000295:instrumentation* / Q000379:methods*
chemical_list: D024042:Collagen Type I
keywords: Active tension; BioMEMS; Differentiation; Myoblasts; Myotubes; Skeletal-muscle-on-a-chip