RT Journal Article
SR Electronic
T1 Single-cell phenotyping of human induced pluripotent stem cells by high-throughput imaging
JF bioRxiv
FD Cold Spring Harbor Laboratory
SP 026955
DO 10.1101/026955
A1 H. Christian Volz
A1 Florian Heigwer
A1 Tatjana Wüest
A1 Marta Galach
A1 Jochen Utikal
A1 Hugo A. Katus
A1 Michael Boutros
YR 2015
UL http://biorxiv.org/content/early/2015/09/16/026955.abstract
AB Single-cell phenotyping promises to yield insights into biological responses in heterogeneous cell populations. We developed a method based on single-cell analysis to phenotype human induced pluripotent stem cells (hIPSC) by high-throughput imaging. Our method uses markers for morphology and pluripotency as well as social features to characterize perturbations using a meta-phenotype based on mapping single cells to distinct phenotypic classes. Analysis of perturbations on a single cell level enhances the applicability of human induced pluripotent stem cells (hIPSC) for screening experiments taking the inherently increased phenotypic variability of these cells into account. We adapted miniaturized culture conditions to allow for the utilization of hIPSC in RNA interference (RNAi) high-throughput screens and single cell phenotyping by image analysis. We identified key regulators of pluripotency in hIPSC masked in a population-averaged analysis and we confirmed several candidate genes (SMG1, TAF1) and assessed their effect on pluripotency.