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

Stem Cell Reports
Thomas F AllisonPeter W Andrews

Abstract

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.

Citations

May 17, 2019·Cancer Biology & Therapy·Ruishuang MaHulun Li
Sep 25, 2019·Nature Cell Biology·Dafne Campigli Di GiammartinoEffie Apostolou
Jun 13, 2020·Nature·Naomi MorisAlfonso Martinez Arias
Oct 7, 2020·Nature Communications·Dylan StavishTariq Enver
Dec 4, 2020·Cell Stem Cell·Masaki KinoshitaAustin Smith
Jul 13, 2021·Frontiers in Cell and Developmental Biology·Daniela Ávila-GonzálezNéstor F Díaz

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Methods Mentioned

BETA
fluorescence activated cell sorting
FACS
flow cytometry
Drop-seq
PCR
single-cell

Software Mentioned

InCell Analyzer 2000

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