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Visualization and quantification of dynamic intercellular coupling in human embryonic stem cells using single cell sonoporation.
- Source :
-
Scientific reports [Sci Rep] 2020 Oct 26; Vol. 10 (1), pp. 18253. Date of Electronic Publication: 2020 Oct 26. - Publication Year :
- 2020
-
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 Ca <superscript>2+</superscript> into single hESCs. We employ live imaging for continuous visualization of intercellular dye transfer and Ca <superscript>2+</superscript> diffusion in hESC colonies. We quantify cell-cell permeability based on dye diffusion using mass transport models. Our results reveal heterogeneous intercellular connectivity and a variety of spatiotemporal characteristics of intercellular Ca <superscript>2+</superscript> waves in hESC colonies induced by sonoporation of single cells.
Details
- Language :
- English
- ISSN :
- 2045-2322
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Scientific reports
- Publication Type :
- Academic Journal
- Accession number :
- 33106521
- Full Text :
- https://doi.org/10.1038/s41598-020-75347-4