title: High-content imaging with micropatterned multiwell plates reveals influence of cell geometry and cytoskeleton on chromatin dynamics.
abstract: Understanding the mechanisms underpinning cellular responses to microenvironmental cues requires tight control not only of the complex milieu of soluble signaling factors, extracellular matrix (ECM) connections and cell-cell contacts within cell culture, but also of the biophysics of human cells. Advances in biomaterial fabrication technologies have recently facilitated detailed examination of cellular biophysics and revealed that constraints on cell geometry arising from the cellular microenvironment influence a wide variety of human cell behaviors. Here, we create an in vitro platform capable of precise and independent control of biochemical and biophysical microenvironmental cues by adapting microcontact printing technology into the format of standard six- to 96-well plates to create MicroContact Printed Well Plates (μCP Well Plates). Automated high-content imaging of human cells seeded on μCP Well Plates revealed tight, highly consistent control of single-cell geometry, cytoskeletal organization, and nuclear elongation. Detailed subcellular imaging of the actin cytoskeleton and chromatin within live human fibroblasts on μCP Well Plates was then used to describe a new relationship between cellular geometry and chromatin dynamics. In summary, the μCP Well Plate platform is an enabling high-content screening technology for human cell biology and cellular engineering efforts that seek to identify key biochemical and biophysical cues in the cellular microenvironment.
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Ty
authors.initials: T
authors.lastname: Harkness
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Mechanical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Jason D
authors.initials: JD
authors.lastname: McNulty
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Ryan
authors.initials: R
authors.lastname: Prestil
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Stephanie K
authors.initials: SK
authors.lastname: Seymour
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Tyler
authors.initials: T
authors.lastname: Klann
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, Yale University, CT, USA.|Systems Biology Institute, Yale University, CT, USA.
authors.forename: Michael
authors.initials: M
authors.lastname: Murrell
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: Randolph S
authors.initials: RS
authors.lastname: Ashton
authors.affiliation: Wisconsin Institute for Discovery, University of Wisconsin-Madison, Madison, WI, USA. ksaha@wisc.edu.|Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA. ksaha@wisc.edu.
authors.forename: Krishanu
authors.initials: K
authors.lastname: Saha
mesh_terms: D008841:Actin Cytoskeleton / Q000648:ultrastructure*; D002450:Cell Communication / Q000235:genetics; D060833:Cellular Microenvironment / Q000235:genetics; D002843:Chromatin / Q000648:ultrastructure*; D005109:Extracellular Matrix / Q000648:ultrastructure; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D008870:Microtubules / Q000648:ultrastructure
chemical_list: D002843:Chromatin
keywords: Actin cytoskeleton; Cell geometry; Chromatin dynamics; High-throughput screening; Microcontact printing