Current cell separation methods rely on density gradient and fluorescent or magnetic labeling. These techniques suffer from poor specificity, use of fluorescent chemicals or antibodies, and high cell loss. Here we present a simple and modular substrate-based separation method that exploits the specificity of cell surface receptors to its ligands for enriching subpopulations of cells. Our strategy employs streptavidin-coated tissue culture plates that display ligands with high specificity towards cell surface receptors. Surfaces displaying the heparin-binding domain of vitronectin (GKKQRFRHRNRKG) are capable of efficiently separating CRISPR-modified heparan sulfate (HS)-deficient human embryonic stem (hES) cells from HS-containing wildtype cells. &gt;75% purity was achieved upon analysis by flow cytometry. Additionally, substrata displaying a small molecule with high specificity towards the aVb3 integrin can separate hES-derived definitive endoderm (DE) cells from undifferentiated hES cells by integrin engagement of DE cells. In summary, our surface-based cell separation strategy represents a novel label-free, high efficiency and recovery method for the enrichment of desired cell subpopulations and is applicable to a wide variety of cell types due to its modular design.

Ligand-specific synthetic surfaces for label-free enrichment of cell subpopulations

256th National Meeting (2018)

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