title: A nanobiosensor for dynamic single cell analysis during microvascular self-organization.
abstract: The formation of microvascular networks plays essential roles in regenerative medicine and tissue engineering. Nevertheless, the self-organization mechanisms underlying the dynamic morphogenic process are poorly understood due to a paucity of effective tools for mapping the spatiotemporal dynamics of single cell behaviors. By establishing a single cell nanobiosensor along with live cell imaging, we perform dynamic single cell analysis of the morphology, displacement, and gene expression during microvascular self-organization. Dynamic single cell analysis reveals that endothelial cells self-organize into subpopulations with specialized phenotypes to form microvascular networks and identifies the involvement of Notch1-Dll4 signaling in regulating the cell subpopulations. The cell phenotype correlates with the initial Dll4 mRNA expression level and each subpopulation displays a unique dynamic Dll4 mRNA expression profile. Pharmacological perturbations and RNA interference of Notch1-Dll4 signaling modulate the cell subpopulations and modify the morphology of the microvascular network. Taken together, a nanobiosensor enables a dynamic single cell analysis approach underscoring the importance of Notch1-Dll4 signaling in microvascular self-organization.
authors.affiliation: Department of Mechanical Engineering, University of Michigan, Ann Arbor, MI 48109, USA.
authors.forename: S
authors.initials: S
authors.lastname: Wang
authors.forename: J
authors.initials: J
authors.lastname: Sun
authors.forename: D D
authors.initials: DD
authors.lastname: Zhang
authors.forename: P K
authors.initials: PK
authors.lastname: Wong
mesh_terms: D048868:Adaptor Proteins, Signal Transducing; D015374:Biosensing Techniques; D002135:Calcium-Binding Proteins; D006046:Gold; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism; D043942:Nanotubes; D009696:Nucleic Acids; D051881:Receptor, Notch1 / Q000378:metabolism; D015398:Signal Transduction; D059010:Single-Cell Analysis
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D002135:Calcium-Binding Proteins; C424126:DLL4 protein, human; D036341:Intercellular Signaling Peptides and Proteins; C497761:NOTCH1 protein, human; D009696:Nucleic Acids; D051881:Receptor, Notch1; D006046:Gold