Acoustic Tweezing Cytometry Induces Rapid Initiation of Human Embryonic Stem Cell Differentiation

Scientific Reports
Tuğba TopalPaul H Krebsbach

Abstract

Mechanical forces play critical roles in influencing human embryonic stem cell (hESC) fate. However, it remains largely uncharacterized how local mechanical forces influence hESC behavior in vitro. Here, we used an ultrasound (US) technique, acoustic tweezing cytometry (ATC), to apply targeted cyclic subcellular forces to hESCs via integrin-bound microbubbles (MBs). We found that ATC-mediated cyclic forces applied for 30 min to hESCs near the edge of a colony induced immediate global responses throughout the colony, suggesting the importance of cell-cell connection in the mechanoresponsiveness of hESCs to ATC-applied forces. ATC application generated increased contractile force, enhanced calcium activity, as well as decreased expression of pluripotency transcription factors Oct4 and Nanog, leading to rapid initiation of hESC differentiation and characteristic epithelial-mesenchymal transition (EMT) events that depend on focal adhesion kinase (FAK) activation and cytoskeleton (CSK) tension. These results reveal a unique, rapid mechanoresponsiveness and community behavior of hESCs to integrin-targeted cyclic forces.

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Citations

Feb 28, 2019·Integrative Biology : Quantitative Biosciences From Nano to Macro·Tuğba TopalPaul H Krebsbach
Nov 14, 2019·Cellular and Molecular Bioengineering·Weiyi Qian, Weiqiang Chen
Dec 2, 2018·Frontiers in Physiology·Francesco BasoliAlberto Rainer
Sep 11, 2021·Scientific Reports·Elnaz Attar Jannesar, Hossein Hamzehpour

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Methods Mentioned

BETA
optical
atomic force microscopy
force measurements
PMA
Protein Assay
immunoprecipitation
sizing
PCR
Assay
Fluorescence

Software Mentioned

excel
Matlab

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