Monitoring developmental force distributions in reconstituted embryonic epithelia

Methods : a Companion to Methods in Enzymology
L PrzybylaV M Weaver

Abstract

The way cells are organized within a tissue dictates how they sense and respond to extracellular signals, as cues are received and interpreted based on expression and organization of receptors, downstream signaling proteins, and transcription factors. Part of this microenvironmental context is the result of forces acting on the cell, including forces from other cells or from the cellular substrate or basement membrane. However, measuring forces exerted on and by cells is difficult, particularly in an in vivo context, and interpreting how forces affect downstream cellular processes poses an even greater challenge. Here, we present a simple method for monitoring and analyzing forces generated from cell collectives. We demonstrate the ability to generate traction force data from human embryonic stem cells grown in large organized epithelial sheets to determine the magnitude and organization of cell-ECM and cell-cell forces within a self-renewing colony. We show that this method can be used to measure forces in a dynamic hESC system and demonstrate the ability to map intracolony protein localization to force organization.

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Citations

Feb 21, 2016·Methods : a Companion to Methods in Enzymology·Nicolas BorghiChristophe Lavelle
Jun 15, 2016·Proceedings of the National Academy of Sciences of the United States of America·Stefan KarpitschkaJacco H Snoeijer
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Sep 19, 2019·Scientific Reports·Seungman ParkYun Chen
Jul 30, 2020·The Journal of Clinical Investigation·Jason J NortheyValerie M Weaver
Sep 12, 2018·Nature Cell Biology·J Matthew BarnesValerie M Weaver
Jan 10, 2020·Biochimica Et Biophysica Acta. Molecular Cell Research·Natalia M GalignianaGraciela Piwien-Pilipuk
Apr 7, 2021·The Journal of Experimental Medicine·Allison P DrainValerie M Weaver
Jun 17, 2021·The Journal of Cell Biology·Vivek K GuptaOvijit Chaudhuri

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