Capturing States of Pluripotency:
How Cell Surface Chemoproteomics Facilitates Rapid Immunophenotyping and Sorting of Pluripotent Stem Cells

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Induction of a pluripotent state in somatic cells through nuclear reprogramming has ushered in a new era of regenerative medicine. However, heterogeneity and varied differentiation potentials among induced pluripotent stem cell (iPSC) lines challenge our ability to identify and isolate functionally defined cell populations for disease modeling, drug discovery, and patient therapies. Thus, there is an urgent need to develop non-genetic methods for isolating more homogeneous, functionally defined iPSCs. To address this, we have applied a highly specific chemoproteomic targeting strategy and high mass accuracy mass spectrometry for the de novo discovery of cell surface exposed N-glycoproteins.

To date, more than 800 cell surface markers on embryonic stem cells (ESCs) and iPSCs have been identified. Utilizing ProteinCenter software, this pluripotent surfaceome map has been compared to the Cell Surface Protein Atlas (D. Bausch-Fluck, in preparation) that contains surfaceome data from more than 80 cell types, to reveal potentially pluripotent-restricted proteins. This new resource has revealed surface markers for sorting functionally distinct stem cell sub-populations and new antibody panels for isolating iPSCs from mixed cultures of reprogrammed cells independent of morphology.

Who should attend?

• Stem cell researchers
• Protein researchers working with cell surface proteins
• Researchers interested in extracting more biologically relevant information from LC/MS protein data

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