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Cellular mechanoadaptation to substrate mechanical properties: contributions of substrate stiffness and thickness to cell stiffness measurements using AFM

DSpace at IIT Bombay

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Title Cellular mechanoadaptation to substrate mechanical properties: contributions of substrate stiffness and thickness to cell stiffness measurements using AFM
 
Creator VICHARE, S
SEN, S
INAMDAR, MM
 
Subject ATOMIC-FORCE MICROSCOPY
INDENTATION
MODULUS
LINEAGE
GROWTH
MODEL
FEEL
 
Description Mechanosensing by adherent cells is usually studied by quantifying cell responses on hydrogels that are covalently linked to a rigid substrate. Atomic force microscopy (AFM) represents a convenient way of characterizing the mechanoadaptation response of adherent cells on hydrogels of varying stiffness and thickness. Since AFM measurements reflect the effective cell stiffness, therefore, in addition to measuring real cytoskeletal alterations across different conditions, these measurements might also be influenced by the geometry and physical properties of the substrate itself. To better understand how the physical attributes of the gel influence AFM stiffness measurements of cells, we have used finite element analysis to simulate the indentation of cells of various spreads resting on hydrogels of varying stiffness and thickness. Consistent with experimental results, our simulation results indicate that for well spread cells, stiffness values are significantly over-estimated when experiments are performed on cells cultured on soft and thin gels. Using parametric studies, we have developed scaling relationships between the effective stiffness probed by AFM and the bulk cell stiffness, taking cell and tip geometry, hydrogel properties, nuclear stiffness and cell contractility into account. Finally, using simulated mechanoadaptation responses, we have demonstrated that a cell stiffening response may arise purely due to the substrate properties. Collectively, our results demonstrate the need to take hydrogel properties into account while estimating cell stiffness using AFM indentation.
 
Publisher ROYAL SOC CHEMISTRY
 
Date 2014-12-28T15:01:24Z
2014-12-28T15:01:24Z
2014
 
Type Article
 
Identifier SOFT MATTER, 10(8)1174-1181
1744-683X
1744-6848
http://dx.doi.org/10.1039/c3sm51786a
http://dspace.library.iitb.ac.in/jspui/handle/100/16819
 
Language English