title: Clonal analysis of individual human embryonic stem cell differentiation patterns in microfluidic cultures.
abstract: Heterogeneity in the clonal outputs of individual human embryonic stem cells (hESCs) confounds analysis of their properties in studies of bulk populations and how to manipulate them for clinical applications. To circumvent this problem we developed a microfluidic device that supports the robust generation of colonies derived from single ESCs. This microfluidic system contains 160 individually addressable chambers equipped for perfusion culture of individual hESCs that could be shown to match the growth rates, marker expression and colony morphologies obtained in conventional cultures. Use of this microfluidic device to analyze the clonal growth kinetics of multiple individual hESCs induced to differentiation revealed variable shifts in the growth rate, area per cell and expression of OCT4 in the progeny of individual hESCs. Interestingly, low OCT4 expression, a slower growth rate and low nuclear to cytoplasmic ratios were found to be correlated responses. This study demonstrates how microfluidic systems can be used to enable large scale live-cell imaging of isolated hESCs exposed to changing culture conditions, to examine how different aspects of their variable responses are correlated.
authors.affiliation: Centre for High-Throughput Biology, University of British Columbia, Vancouver, BC, Canada.|Department of Chemical and Biological Engineering, University of British Columbia, Vancouver, BC, Canada.|Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Darek J
authors.initials: DJ
authors.lastname: Sikorski
authors.affiliation: Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Nicolas J
authors.initials: NJ
authors.lastname: Caron
authors.affiliation: Centre for High-Throughput Biology, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Michael
authors.initials: M
authors.lastname: VanInsberghe
authors.affiliation: Centre for High-Throughput Biology, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Hans
authors.initials: H
authors.lastname: Zahn
authors.affiliation: Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, BC, Canada.|Department of Medical Genetics, University of British Columbia, Vancouver, BC, Canada.
authors.forename: Connie J
authors.initials: CJ
authors.lastname: Eaves
authors.affiliation: Department of Chemical and Biological Engineering, University of British Columbia, Vancouver, BC, Canada.|Michael Smith Laboratories, University of British Columbia, Vancouver, BC, Canada.
authors.forename: James M
authors.initials: JM
authors.lastname: Piret
authors.affiliation: Centre for High-Throughput Biology, University of British Columbia, Vancouver, BC, Canada. chansen@phas.ubc.ca.|Department of Physics and Astronomy, University of British Columbia, Vancouver, BC, Canada. chansen@phas.ubc.ca.
authors.forename: Carl L
authors.initials: CL
authors.lastname: Hansen
mesh_terms: D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002454:Cell Differentiation / Q000235:genetics; D019070:Cell Lineage; D049109:Cell Proliferation / Q000235:genetics; D005434:Flow Cytometry; D018740:Genetic Heterogeneity; D000066449:Human Embryonic Stem Cells / Q000166:cytology*; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation / Q000379:methods*; D039904:Pluripotent Stem Cells / Q000166:cytology*
keywords: Clonal analysis; Heterogeneity; Human embryonic stem cell (hESC); Microfluidic cell culture