Abstract
Morphogenesis involves interactions of asymmetric cell populations to form complex multicellular patterns and structures comprised of distinct cell types. However, current methods to model morphogenic events lack control over cell-type co-emergence and offer little capability to selectively perturb specific cell subpopulations. Our in vitro system interrogates cell-cell interactions and multicellular organization within human induced pluripotent stem cell (hiPSC) colonies. We examined effects of induced mosaic knockdown of molecular regulators of cortical tension (ROCK1) and cell-cell adhesion (CDH1) with CRISPR interference. Mosaic knockdown of ROCK1 or CDH1 resulted in differential patterning within hiPSC colonies due to cellular self-organization, while retaining an epithelial pluripotent phenotype. Knockdown induction stimulates a transient wave of differential gene expression within the mixed populations that stabilized in coordination with observed self-organization. Mosaic patterning enables genetic interrogation of emergent multicellular properties, which can facilitate better understanding of the molecular pathways that regulate symmetry-breaking during morphogenesis.
Highlights
Morphogenic tissue development requires the robust coordination of biochemical and biophysical signaling cues to dictate cell-cell communication, multicellular organization, and cell fate determination. (Burdsal et al, 1993; Leckband et al, 2011; Montero and Heisenberg, 2004)
Protein KD followed a similar trend where KD populations compared to untreated controls resulted in
In this study we examined the effect of inducing specific genetic KD in subpopulations of human induced pluripotent stem cell (hiPSC) within an otherwise homogeneous population of pluripotent cells
Summary
Morphogenic tissue development requires the robust coordination of biochemical and biophysical signaling cues to dictate cell-cell communication, multicellular organization, and cell fate determination. (Burdsal et al, 1993; Leckband et al, 2011; Montero and Heisenberg, 2004). Morphogenic tissue development requires the robust coordination of biochemical and biophysical signaling cues to dictate cell-cell communication, multicellular organization, and cell fate determination. A hallmark of morphogenesis is the asymmetric co-emergence of distinct cell populations that selforganize to form developmental patterns, multicellular structures, and functional tissues and organs (Bronner, 2016; Lancaster and Knoblich, 2014; Sasai, 2013). During gastrulation, the blastocyst transitions from a relatively homogeneous population of pluripotent cells to a spatially organized, multicellular composition of distinct progenitor cells. To study morphogenesis, it is essential to promote the coincident development of analogous heterogeneous populations in vitro. Human pluripotent stem cells (hPSCs) provide an unlimited source of cells that can
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