Abstract
SummaryNaive and primed human pluripotent stem cells (hPSC) provide valuable models to study cellular and molecular developmental processes. The lack of detailed information about cell-surface protein expression in these two pluripotent cell types prevents an understanding of how the cells communicate and interact with their microenvironments. Here, we used plasma membrane profiling to directly measure cell-surface protein expression in naive and primed hPSC. This unbiased approach quantified over 1,700 plasma membrane proteins, including those involved in cell adhesion, signaling, and cell interactions. Notably, multiple cytokine receptors upstream of JAK-STAT signaling were more abundant in naive hPSC. In addition, functional experiments showed that FOLR1 and SUSD2 proteins are highly expressed at the cell surface in naive hPSC but are not required to establish human naive pluripotency. This study provides a comprehensive stem cell proteomic resource that uncovers differences in signaling pathway activity and has identified new markers to define human pluripotent states.
Highlights
Recent reports have identified conditions that can stabilize human pluripotent stem cells in different states that hold distinct molecular and functional properties (Guo et al, 2017; Takashima et al, 2014; Theunissen et al, 2014)
Our unbiased approach identified over 1,700 plasma membrane proteins, including those involved in cell adhesion, signaling, and cell interactions, thereby providing a comprehensive stem cell proteomic resource
We examined the expression of cell-surface proteins that are upstream of transforming growth factor b (TGF-b) signaling, as this pathway is required for primed human pluripotent stem cells (hPSC) selfrenewal (Vallier et al, 2004)
Summary
Recent reports have identified conditions that can stabilize human pluripotent stem cells (hPSC) in different states that hold distinct molecular and functional properties (Guo et al, 2017; Takashima et al, 2014; Theunissen et al, 2014). At the two ends of the pluripotency spectrum are naive hPSC that recapitulate the pre-implantation human epiblast, and conventionally grown primed hPSC that more closely align to the post-implantation epiblast (Dong et al, 2019). These cell types can provide informative models to study the earliest stages of human development and the control of pluripotent cell identity (Davidson et al, 2015; Weinberger et al, 2016). Strategies for enriching or eliminating target cell types within a population often rely on the known expression patterns of cell-surface markers and mapping surface proteomes is, a prerequisite for applying these approaches in naive and primed hPSC (Ben-David et al, 2013; Choo et al, 2008; Tang et al, 2011)
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