title: Integrated Colony Imaging, Analysis, and Selection Device for Regenerative Medicine.
abstract: Stem and progenitor cells derived from human tissues are being developed as cell sources for cell-based assays and therapies. However, tissue-derived stem and progenitor cells are heterogeneous. Differences in observed clones of stem cells likely reflect important aspects of the underlying state of the source cells, as well as future potency for cell therapies. This paper describes a colony analysis and picking device that provides quantitative analysis of heterogeneous cell populations and precise tools for cell picking for research or biomanufacturing applications. We describe an integrated robotic system that enables image acquisition and automated image analysis to be coupled with rapid automated selection of individual colonies in adherent cell cultures. Other automated systems have demonstrated feasibility with picking from semisolid media or off feeder layers. We demonstrate the capability to pick adherent bone-derived stem cells from tissue culture plastic. Cells are efficiently picked from a target site and transferred to a recipient well plate. Cells demonstrate viability and adherence and maintain biologic potential for surface markers CD73 and CD90 based on phase contrast and fluorescence imaging 6 days after transfer. Methods developed here can be applied to the study of other stem cell types and automated culture of cells.
authors.affiliation: 1 Department of Biomedical Engineering, Cleveland Clinic, Cleveland, OH, USA.|2 Department of Biomedical Engineering, Case Western Reserve University, Cleveland, OH, USA.
authors.forename: Edward
authors.initials: E
authors.lastname: Kwee
authors.affiliation: 3 eehscience LLC, Pickerington, OH, USA.
authors.forename: Edward E
authors.initials: EE
authors.lastname: Herderick
authors.affiliation: 4 Parker Hannifin Electromechanical, Irwin, PA, USA.
authors.forename: Thomas
authors.initials: T
authors.lastname: Adams
authors.affiliation: 4 Parker Hannifin Electromechanical, Irwin, PA, USA.
authors.forename: James
authors.initials: J
authors.lastname: Dunn
authors.affiliation: 4 Parker Hannifin Electromechanical, Irwin, PA, USA.
authors.forename: Robert
authors.initials: R
authors.lastname: Germanowski
authors.affiliation: 4 Parker Hannifin Electromechanical, Irwin, PA, USA.
authors.forename: Frank
authors.initials: F
authors.lastname: Krakosh
authors.affiliation: 1 Department of Biomedical Engineering, Cleveland Clinic, Cleveland, OH, USA.
authors.forename: Cynthia
authors.initials: C
authors.lastname: Boehm
authors.affiliation: 4 Parker Hannifin Electromechanical, Irwin, PA, USA.
authors.forename: James
authors.initials: J
authors.lastname: Monnich
authors.affiliation: 5 Department of Biomedical Informatics, The Ohio State University, Columbus, OH, USA.
authors.forename: Kimerly
authors.initials: K
authors.lastname: Powell
authors.affiliation: 1 Department of Biomedical Engineering, Cleveland Clinic, Cleveland, OH, USA.
authors.forename: George
authors.initials: G
authors.lastname: Muschler
mesh_terms: D057205:Automation, Laboratory / Q000379:methods; D003584:Cytological Techniques / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D007091:Image Processing, Computer-Assisted / Q000295:instrumentation / Q000379:methods; D061848:Optical Imaging / Q000295:instrumentation / Q000379:methods*; D044968:Regenerative Medicine / Q000295:instrumentation / Q000379:methods*; D012371:Robotics / Q000379:methods; D013234:Stem Cells / Q000502:physiology*
keywords: cell picking; image analysis; microscopy; robotics and instrumentation; stem and progenitor cells