Visualization and quantification of dynamic intercellular coupling in human embryonic stem cells using single cell sonoporation.

ABSTRACT: Gap junctions (GJs), which are proteinaceous channels, couple adjacent cells by permitting direct exchange of intracellular molecules with low molecular weights. GJ intercellular communication (GJIC) plays a critical role in regulating behaviors of human embryonic stem cells (hESCs), affecting their proliferation and differentiation. Here we report a novel use of sonoporation that enables single cell intracellular dye loading and dynamic visualization/quantification of GJIC in hESC colonies. By applying a short ultrasound pulse to excite single microbubbles tethered to cell membranes, a transient pore on the cell membrane (sonoporation) is generated which allows intracellular loading of dye molecules and influx of Ca²⁺ into single hESCs. We employ live imaging for continuous vis

SUBMITTER: Fan Z 

PROVIDER: S-EPMC7589565 | biostudies-literature | 2020 Oct

REPOSITORIES: biostudies-literature