title: High throughput assembly of spatially controlled 3D cell clusters on a micro/nanoplatform.
abstract: Guided assembly of microscale tissue subunits (i.e. 3D cell clusters/aggregates) has found applications in cell therapy/tissue engineering, cell and developmental biology, and drug discovery. As cluster size and geometry are known to influence cellular responses, the ability to spatially control cluster formation in a high throughput manner could be advantageous for many biomedical applications. In this work, a micro- and nanofabricated platform was developed for this purpose, consisting of a soft-lithographically fabricated array of through-thickness microwells structurally bonded to a sheet of electrospun fibers. The microwells and fibers were manufactured from several polymers of biomedical interest. Human hepatocytes were used as model cells to demonstrate the ability of the platform to allow controlled cluster formation. In addition, the ability of the device to support studies on semi-controlled heterotypic interactions was demonstrated by co-culturing hepatocytes and fibroblasts. Preliminary experiments with other cells of interest (pancreatic cells, embryonic stem cells, and cardiomyocytes) were also conducted. Our platform possesses several advantages over previously developed microwell arrays: a more in vivo-like topographical stimulation of cells; better nutrient/waste exchange through the underlying nanofiber mat; and easy integration into standard two-chamber cell culture well systems.
authors.affiliation: Department of Biomedical Engineering, The Ohio State University, 270 Bevis Hall, 1080 Carmack Rd., Columbus, OH 43210, USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Gallego-Perez
authors.forename: Natalia
authors.initials: N
authors.lastname: Higuita-Castro
authors.forename: Sadhana
authors.initials: S
authors.lastname: Sharma
authors.forename: Rashmeet K
authors.initials: RK
authors.lastname: Reen
authors.forename: Andre F
authors.initials: AF
authors.lastname: Palmer
authors.forename: Keith J
authors.initials: KJ
authors.lastname: Gooch
authors.forename: L James
authors.initials: LJ
authors.lastname: Lee
authors.forename: John J
authors.initials: JJ
authors.lastname: Lannutti
authors.forename: Derek J
authors.initials: DJ
authors.lastname: Hansford
mesh_terms: D000818:Animals; D002450:Cell Communication / Q000502:physiology; D002460:Cell Line; D002469:Cell Separation / Q000295:instrumentation*; D018920:Coculture Techniques / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D005347:Fibroblasts / Q000166:cytology / Q000502:physiology*; D022781:Hepatocytes / Q000166:cytology / Q000502:physiology*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D036103:Nanotechnology / Q000295:instrumentation*