Mechanosensitivity to extracellular mechanical signals is central to many developmental, physiological, and pathological processes, affecting cell functions including growth, migration, differentiation, and apoptosis. Understanding the molecular mechanisms underlying mechanotransduction process rely on tools capable of applying controlled mechanical forces to cells to elicit and assess cellular responses. The goal of this research is to develop a novel ultrasound-based technology, acoustic tweezing cytometry (ATC), as a powerful cell mechanics and mechanobiology tool. We will perform systematic and comprehensive studies to develop innovative ATC platform and characterize subcellular force generation in ATC for mechanical regulation of cells, which will have broad impact on many practical applications as well as scientific investigations. In this research, we will develop and demonstrate the utility of ATC as a novel and practical strategy for stem cell applications, specifically to enable novel advances in human pluripotent stem cell (hPSC) maintenance and understanding of mechanobiology of hPSCs. Capable of replicating themselves while retaining the ability to give rise to any type of specialized cells, hPSCs provide promising sources for disease modeling, drug screenings, and future cell-based therapeutics to treat degenerative diseases such as diabetes mellitus and spinal cord injury. However, controlling hPSC growth remains challenging because present methods to clonally grow hPSCs are inefficient and poorly defined for genetic manipulation and therapeutic purposes. hPSCs are vulnerable to apoptosis upon cellular detachment and dissociation, with a cloning efficiency of dissociated single hPSCs generally <1%. Therefore, we propose the following specific aims in this research: 1) to develop an innovative ATC technology platform for applying spatiotemporally controlled subcellular mechanical forces;2) to determine the effects of ATC on the survival and cloning efficiency of hPSCs;and 3) to reveal the mechanisms of ATC stimulation for improving survival and cloning efficiency of hPSCs.

Public Health Relevance

Mechnotransduction is central to many developmental, physiological, and pathological processes and studying mechanotransduction relies on tools for applying mechanical forces to cells to elicit cellular responses. Human pluripotent stem cells (hPSCs) are invaluable tools for studying tissue formation and promising resource for regenerative medicine. This research will develop a novel technology, acoustic tweezing cytometry, as a powerful tool for cell mechanics and mechanobiology studies, and will specifically exploit its application for improving survival threshold and cloning efficiency of (hPSCs), a major challenge limiting their wide applications.

Agency
National Institute of Health (NIH)
Institute
National Institute of Biomedical Imaging and Bioengineering (NIBIB)
Type
Research Project (R01)
Project #
1R01EB019436-01A1
Application #
8896236
Study Section
Medical Imaging Study Section (MEDI)
Program Officer
Conroy, Richard
Project Start
2015-04-15
Project End
2019-01-31
Budget Start
2015-04-15
Budget End
2016-01-31
Support Year
1
Fiscal Year
2015
Total Cost
$496,591
Indirect Cost
$167,612
Name
University of Michigan Ann Arbor
Department
Biomedical Engineering
Type
Schools of Engineering
DUNS #
073133571
City
Ann Arbor
State
MI
Country
United States
Zip Code
48109
Shao, Yue; Sang, Jianming; Fu, Jianping (2015) On human pluripotent stem cell control: The rise of 3D bioengineering and mechanobiology. Biomaterials 52:26-43