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Characterization of Human Embryonic Stem Cell (hESC)-derived Teratomas via Single Cell RNA-Sequencing

  • Author(s): Yusupova, Marianna
  • Advisor(s): Mali, Prashant
  • et al.
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Abstract

Although the advent of human pluripotent stem cell (hPSC)-based 2D and 3D models has enabled great strides in our understanding of human development and disease, the limitations of these models, particularly the absence of cellular interactions across different lineages as well as vascular networks, make it difficult to accurately recapitulate molecular and physiological processes in vitro. Towards addressing this, we have characterized human embryonic stem cell (hESC)-derived teratomas in order to investigate their potential as alternative 3D modeling systems. As teratomas are naturally comprised of cells representing each of the three germ layers (endoderm, mesoderm, ectoderm) and vascularized, we believe such a model may enable recapitulation of cell-cell interactions and developmental processes otherwise difficult to study in a context relevant to human biology. As an initial step, we employed single cell RNA-sequencing (SC RNA-seq) methods for characterization of the cellular composition, diversity, and consistency of cell populations across hESC-derived teratomas. We found that as expected, cell lineages derived from each of the three germ layers (endoderm, mesoderm, ectoderm), as well as both early progenitor and more mature differentiated cell types, were present, along with different cell states. Additionally, we found that methodologies used to process teratomas play a critical role in scRNA-seq data interpretation. However, despite intrinsic teratoma heterogeneity, teratoma samples in general showed close alignment and representation across clusters, indicating a level of predictability to this system. Taken together, the results indicate that teratomas hold potential as a novel alternative modeling system for the study and recapitulation of normal and diseased multi-lineage human development.

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This item is under embargo until June 22, 2020.