PT  - JOURNAL ARTICLE
AU  - H. Christian Volz
AU  - Florian Heigwer
AU  - Tatjana Wüest
AU  - Marta Galach
AU  - Jochen Utikal
AU  - Hugo A. Katus
AU  - Michael Boutros
TI  - Single-cell phenotyping of human induced pluripotent stem cells by high-throughput imaging
AID  - 10.1101/026955
DP  - 2015 Jan 01
TA  - bioRxiv
PG  - 026955
4099  - http://biorxiv.org/content/early/2015/09/16/026955.short
4100  - http://biorxiv.org/content/early/2015/09/16/026955.full
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.