Deterministic Single Cell Encapsulation in Asymmetric Microenvironments to Direct Cell Polarity

Various signals in tissue microenvironments are often unevenly distributed around cells. Cellular responses to asymmetric cell-matrix adhesion in a 3D space remain generally unclear and are to be studied at the single-cell resolution. Here, the authors developed a droplet-based microfluidic approach to manufacture a pure population of single cells in a microscale layer of compartmentalized 3D hydrogel matrices with a tunable spatial presentation of ligands at the subcellular level. Cells elongate with an asymmetric presentation of the integrin adhesion ligand Arg-Gly-Asp (RGD), while cells expand isotropically with a symmetric presentation of RGD. Membrane tension is higher on the side of single cells interacting with RGD than on the side without RGD. Finite element analysis shows that a non-uniform isotropic cell volume expansion model is sufficient to recapitulate the experimental results. At a longer timescale, asymmetric ligand presentation commits mesenchymal stem cells to the osteogenic lineage. Cdc42 is an essential mediator of cell polarization and lineage specification in response to asymmetric cell-matrix adhesion. This study highlights the utility of precisely controlling 3D ligand presentation around single cells to direct cell polarity for regenerative engineering and medicine.

Automated summary

The researchers developed a microfluidic approach to create single cells in compartmentalized 3D hydrogel matrices with tunable ligand presentation. They found that cells elongate with asymmetric presentation of integrin adhesion ligands, while they expand isotropically with symmetric presentation. Furthermore, they discovered that asymmetric ligand presentation influences the lineage commitment of stem cells. This study demonstrates the potential of precisely controlling 3D ligand presentation to direct cell polarity in regenerative engineering and medicine.