Identification and Single-Cell Functional Characterization of an Endodermally Biased Pluripotent Substate in Human Embryonic Stem Cells

• Published in: Stem cell reports 2018-06, Vol.10 (6), p.1895-1907
• Main Authors: Allison, Thomas F., Smith, Andrew J.H., Anastassiadis, Konstantinos, Sloane-Stanley, Jackie, Biga, Veronica, Stavish, Dylan, Hackland, James, Sabri, Shan, Langerman, Justin, Jones, Mark, Plath, Kathrin, Coca, Daniel, Barbaric, Ivana, Gokhale, Paul, Andrews, Peter W.
• Format: Article
• Language: English
• Subjects: Biomarkers; Cell Differentiation - genetics; Cell Self Renewal; differentiation bias; Endoderm - cytology; GATA6; GATA6 Transcription Factor - genetics; GATA6 Transcription Factor - metabolism; Gene Expression Profiling; Genes, Reporter; human embryonic stem cell heterogeneity; Human Embryonic Stem Cells - cytology; Human Embryonic Stem Cells - metabolism; Humans; Immunophenotyping; lineage priming; Single-Cell Analysis - methods
• ISSN: 2213-6711; 2213-6711
• DOI: 10.1016/j.stemcr.2018.04.015

Human embryonic stem cells (hESCs) display substantial heterogeneity in gene expression, implying the existence of discrete substates within the stem cell compartment. To determine whether these substates impact fate decisions of hESCs we used a GFP reporter line to investigate the properties of fractions of putative undifferentiated cells defined by their differential expression of the endoderm transcription factor, GATA6, together with the hESC surface marker, SSEA3. By single-cell cloning, we confirmed that substates characterized by expression of GATA6 and SSEA3 include pluripotent stem cells capable of long-term self-renewal. When clonal stem cell colonies were formed from GATA6-positive and GATA6-negative cells, more of those derived from GATA6-positive cells contained spontaneously differentiated endoderm cells than similar colonies derived from the GATA6-negative cells. We characterized these discrete cellular states using single-cell transcriptomic analysis, identifying a potential role for SOX17 in the establishment of the endoderm-biased stem cell state. [Display omitted] •Subsets of hESCs can co-express pluripotency-associated and lineage-specific genes•hESCs co-expressing GATA6 are capable of long-term self-renewal•Single GATA6-expressing hESCs regenerate GATA6-negative cells•GATA6-expressing hESCs are biased in their propensity for differentiation Human embryonic stem cells have the capacity to turn into any cell type within the adult. Peter Andrews and colleagues have shown that subtle differences between individual cells can functionally bias the resulting cell type that is produced. Generating and purifying these biased cells may therefore improve the derivation of medically relevant cell types.