A qPCR ScoreCard quantifies the differentiation potential of human pluripotent stem cells.
Research on human pluripotent stem cells has been hampered by the lack of a standardized, quantitative, scalable assay of pluripotency. We previously described an assay called ScoreCard that used gene expression signatures to quantify differentiation efficiency. Here we report an improved version of the assay based on qPCR that enables faster, more quantitative assessment of functional pluripotency. We provide an in-depth characterization of the revised signature panel (commercially available as the TaqMan hPSC Scorecard Assay) through embryoid body and directed differentiation experiments as well as a detailed comparison to the teratoma assay. We further show that the improved ScoreCard enables a wider range of applications, such as screening of small molecules, genetic perturbations and assessment of culture conditions. Our approach can be extended beyond stem cell applications to characterize and assess the utility of other cell types and lineages.
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.
Alexander M
AM
Tsankov
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.
Veronika
V
Akopian
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.
Ramona
R
Pop
Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.
Sundari
S
Chetty
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.
Casey A
CA
Gifford
Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.
Laurence
L
Daheron
Department of Pathology and Cell Biology, Columbia University Medical Center, New York, New York, USA.|Department of Pathology, Icahn School of Medicine at Mount Sinai, New York, New York, USA.|Department of Neuroscience, Icahn School of Medicine at Mount Sinai, New York, New York, USA.
Nadejda M
NM
Tsankova
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.|Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.|Department of Stem Cell and Regenerative Biology, Harvard University, Cambridge, Massachusetts, USA.
Alexander
A
Meissner
D000818:Animals; D002454:Cell Differentiation / Q000235:genetics*; D019295:Computational Biology / Q000379:methods*; D058732:Embryoid Bodies / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D006801:Humans; D051379:Mice; D009374:Neoplasms, Experimental / Q000473:pathology; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000502:physiology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D013724:Teratoma / Q000473:pathology