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Abstract Cells in living tissues are exposed to substantial mechanical forces and constraints imposed by neighboring cells, the extracellular matrix, and external factors. Mechanical forces and physical confinement can drive various cellular responses, including changes in gene expression, cell growth, differentiation, and migration, all of which have important implications in physiological and pathological processes, such as immune cell migration or cancer metastasis. Previous studies have shown that nuclear deformation induced by 3D confinement promotes cell contractility but can also cause DNA damage and changes in chromatin organization, thereby motivating further studies in nuclear mechanobiology. In this protocol, we present a custom‐developed, easy‐to‐use, robust, and low‐cost approach to induce precisely defined physical confinement on cells using agarose pads with micropillars and externally applied weights. We validated the device by confirming nuclear deformation, changes in nuclear area, and cell viability after confinement. The device is suitable for short‐ and long‐term confinement studies and compatible with imaging of both live and fixed samples, thus presenting a versatile approach to studying the impact of 3D cell confinement and nuclear deformation on cellular function. This article contains detailed protocols for the fabrication and use of the confinement device, including live cell imaging and labeling of fixed cells for subsequent analysis. These protocols can be amended for specific applications. © 2023 Wiley Periodicals LLC. Basic Protocol 1: Design and fabrication of the confinement device wafer Basic Protocol 2: Cell confinement assay Support Protocol 1: Fixation and staining of cells after confinement Support protocol 2: Live/dead staining of cells during confinement
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Frontiers in mechanobiology and mechanomedicine
Abstract This perspective derives from the presentations and discussions on mechanobiology at the 2025 Cellular and Molecular Bioengineering Conference in San Diego. Mechanobiological processes play critical roles in tissue development, regeneration, and disease progression. Recent advances in engineering, biology, and medicine have enabled the translation of mechanobiology discoveries into clinical practice, giving rise to the emerging field of mechanomedicine. The development and application of engineering technology and tools have provided new insights into how mechanical cues regulate immune cell response, stem cell differentiation, cell migration, and cell metabolism. In this perspective, we highlight exciting discoveries and innovative tools in mechanobiology research, and discuss challenges that must be overcome to truly bridge the gap between mechanobiology and mechanomedicine. Graphical Abstract
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- Award ID(s):
- 2145756
- PAR ID:
- 10657544
- Publisher / Repository:
- Springer Nature
- Date Published:
- Journal Name:
- Med-X
- Volume:
- 3
- Issue:
- 1
- ISSN:
- 2097-440X
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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- Free Publicly Accessible Full Text
-
Accepted Manuscript
- Journal Article:
- https://doi.org/10.1007/s44258-025-00071-5
