Monitoring developmental force distributions in reconstituted embryonic epithelia.
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.
Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California, San Francisco (UCSF), San Francisco, CA, USA.
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Przybyla
Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California, San Francisco (UCSF), San Francisco, CA, USA.
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Lakins
Institute for Bioengineering of Catalonia (IBEC), Baldiri Reixac 15-21, 08028 Barcelona, Spain.
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Sunyer
Institute for Bioengineering of Catalonia (IBEC), Baldiri Reixac 15-21, 08028 Barcelona, Spain; Facultat de Medicina, Universitat de Barcelona, 08036 Barcelona, Spain; Institució Catalana de Recerca i Estudis Avançats, 08010 Barcelona, Spain.
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Trepat
Center for Bioengineering and Tissue Regeneration, Department of Surgery, University of California, San Francisco (UCSF), San Francisco, CA, USA; Department of Anatomy and Department of Bioengineering and Therapeutic Sciences, Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research and Helen Diller Family Comprehensive Cancer Center, UCSF, San Francisco, CA, USA. Electronic address: Valerie.Weaver@ucsf.edu.
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Weaver
D002454:Cell Differentiation; D002465:Cell Movement; D002478:Cells, Cultured; D000066449:Human Embryonic Stem Cells / Q000502:physiology*; D006801:Humans; D040542:Mechanotransduction, Cellular; D018625:Microscopy, Atomic Force
Epiblast; Human embryonic stem cells; Mechanotransduction; Monolayer stress microscopy; Self-organization; Traction force