title: A gel-free 3D microfluidic cell culture system.
abstract: 3D microfluidic cell culture systems offer a biologically relevant model to conduct micro-scale mammalian cell-based research and applications. Various natural and synthetic hydrogels have been successfully incorporated into microfluidic systems to support mammalian cells in 3D. However, embedment of cells in hydrogels introduces operational complexity, potentially hinders mass transfer, and is not suitable for establishing cell-dense, ECM-poor constructs. We present here a gel-free method for seeding and culturing mammalian cells three-dimensionally in a microfluidic channel. A combination of transient inter-cellular polymeric linker and micro-fabricated pillar arrays was used for the in situ formation and immobilization of 3D multi-cellular aggregates in a microfluidic channel. 3D cellular constructs formed this way are relieved of hydrogel embedment for cellular support. Two mammalian cell lines (A549 and C3A) and a primary mammalian cell (bone marrow mesenchymal stem cells) were cultured in the gel-free 3D microfluidic cell culture system. The cells displayed 3D cellular morphology, cellular functions and differentiation capability, affirming the versatility of the system as a 3D cell perfusion culture platform for anchorage-dependent mammalian cells.
authors.affiliation: Institute of Bioengineering and Nanotechnology, A*STAR, The Nanos, #04-01, 31 Biopolis Way, Singapore 138669, Singapore.
authors.forename: Siew-Min
authors.initials: SM
authors.lastname: Ong
authors.forename: Chi
authors.initials: C
authors.lastname: Zhang
authors.forename: Yi-Chin
authors.initials: YC
authors.lastname: Toh
authors.forename: So Hyun
authors.initials: SH
authors.lastname: Kim
authors.forename: Hsien Loong
authors.initials: HL
authors.lastname: Foo
authors.forename: Choon Hong
authors.initials: CH
authors.lastname: Tan
authors.forename: Danny
authors.initials: D
authors.lastname: van Noort
authors.forename: Sungsu
authors.initials: S
authors.lastname: Park
authors.forename: Hanry
authors.initials: H
authors.lastname: Yu
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D002449:Cell Aggregation; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002460:Cell Line; D002470:Cell Survival; D002478:Cells, Cultured; D006801:Humans; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D044085:Microfluidics / Q000295:instrumentation / Q000379:methods*; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D051381:Rats
chemical_list: D000199:Actins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate