Adipocyte stiffness increases with accumulation of lipid droplets.

Adipogenesis and increase in fat tissue mass are mechanosensitive processes and hence should be influenced by the mechanical properties of adipocytes. We evaluated subcellular effective stiffnesses of adipocytes using atomic force microscopy (AFM) and interferometric phase microscopy (IPM), and we verified the empirical results using finite element (FE) simulations. In the AFM studies, we found that the mean ratio of stiffnesses of the lipid droplets (LDs) over the nucleus was 0.83 ± 0.14, from which we further evaluated the ratios of LDs over cytoplasm stiffness, as being in the range of 2.5 to 8.3. These stiffness ratios, indicating that LDs are stiffer than cytoplasm, were verified by means of FE modeling, which simulated the AFM experiments, and provided good agreement between empirical and model-predicted structural behavior. In the IPM studies, we found that LDs mechanically distort their intracellular environment, which again indicated that LDs are mechanically stiffer than the surrounding cytoplasm. Combining these empirical and simulation data together, we provide in this study evidence that adipocytes stiffen with differentiation as a result of accumulation of LDs. Our results are relevant to research of adipose-related diseases, particularly overweight and obesity, from a mechanobiology and cellular mechanics perspectives.

[1]  Gabriel Popescu,et al.  Measurement of red blood cell mechanics during morphological changes , 2010, Proceedings of the National Academy of Sciences.

[2]  Amit Gefen,et al.  Mechanotransduction in adipocytes. , 2012, Journal of biomechanics.

[3]  M. Wabitsch,et al.  Reorganization of the nuclear lamina and cytoskeleton in adipogenesis , 2011, Histochemistry and Cell Biology.

[4]  Pinhas Girshovitz,et al.  Generalized cell morphological parameters based on interferometric phase microscopy and their application to cell life cycle characterization , 2012, Biomedical optics express.

[5]  A. Gefen,et al.  Stochastic Modeling of Adipogenesis in 3T3-L1 Cultures to Determine Probabilities of Events in the Cell’s Life Cycle , 2011, Annals of Biomedical Engineering.

[6]  Gabriel Popescu,et al.  Measurement of the nonlinear elasticity of red blood cell membranes. , 2011, Physical review. E, Statistical, nonlinear, and soft matter physics.

[7]  Natan T. Shaked,et al.  Whole-cell-analysis of live cardiomyocytes using wide-field interferometric phase microscopy , 2010, Biomedical optics express.

[8]  A. Gefen,et al.  Evaluating the effective shear modulus of the cytoplasm in cultured myoblasts subjected to compression using an inverse finite element method. , 2011, Journal of the mechanical behavior of biomedical materials.

[9]  Lay Poh Tan,et al.  Mechanical behavior of human mesenchymal stem cells during adipogenic and osteogenic differentiation. , 2010, Biochemical and biophysical research communications.

[10]  A. Gefen,et al.  The Biomechanics of Fat: From Tissue to a Cell Scale , 2016 .

[11]  S Levin,et al.  Membrane fluctuations in erythrocytes are linked to MgATP-dependent dynamic assembly of the membrane skeleton. , 1991, Biophysical journal.

[12]  Gabriel Popescu,et al.  Optical imaging of cell mass and growth dynamics. , 2008, American journal of physiology. Cell physiology.

[13]  R. Skalak,et al.  Cytoplasmic rheology of passive neutrophils. , 1991, Biorheology.

[14]  C. S. Chen,et al.  Demonstration of mechanical connections between integrins, cytoskeletal filaments, and nucleoplasm that stabilize nuclear structure. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[15]  T. Farley The role of government in preventing excess calorie consumption: the example of New York City. , 2012, JAMA.

[16]  Elizabeth G Loboa,et al.  Cytoskeletal and focal adhesion influences on mesenchymal stem cell shape, mechanical properties, and differentiation down osteogenic, adipogenic, and chondrogenic pathways. , 2012, Tissue engineering. Part B, Reviews.

[17]  Amit Gefen,et al.  Quantitative monitoring of lipid accumulation over time in cultured adipocytes as function of culture conditions: toward controlled adipose tissue engineering. , 2010, Tissue engineering. Part C, Methods.

[18]  Pinhas Girshovitz,et al.  Optical‐mechanical signatures of cancer cells based on fluctuation profiles measured by interferometry , 2014, Journal of biophotonics.

[19]  Cees W J Oomens,et al.  Predicting local cell deformations in engineered tissue constructs: a multilevel finite element approach. , 2002, Journal of biomechanical engineering.

[20]  J. Mertz Introduction to Optical Microscopy , 2009 .

[21]  A. Gefen,et al.  Confocal microscopy-based three-dimensional cell-specific modeling for large deformation analyses in cellular mechanics. , 2010, Journal of biomechanics.

[22]  É. Szabó,et al.  Cytoskeletal Disassembly and Cell Rounding Promotes Adipogenesis from ES Cells , 2010, Stem Cell Reviews and Reports.

[23]  Gabriel Popescu,et al.  Imaging red blood cell dynamics by quantitative phase microscopy. , 2008, Blood cells, molecules & diseases.

[24]  Suliana Manley,et al.  Optical measurement of cell membrane tension. , 2006, Physical review letters.

[25]  N. Caille,et al.  Contribution of the nucleus to the mechanical properties of endothelial cells. , 2002, Journal of biomechanics.

[26]  David R. Kaeli,et al.  Accelerating mesh-based Monte Carlo method on modern CPU architectures , 2012, Biomedical optics express.

[27]  G. G. Bilodeau,et al.  Regular Pyramid Punch Problem , 1992 .

[28]  Ben Fabry,et al.  Cytoskeletal remodelling and slow dynamics in the living cell , 2005, Nature materials.

[29]  J. Lammerding,et al.  Nuclear Shape, Mechanics, and Mechanotransduction , 2008, Circulation research.

[30]  A. Bstr Association of all-cause mortality with overweight and obesity using standard body mass index categories. A systematic review and meta-analysis , 2013, BDJ.

[31]  M. Lane,et al.  Adipose Development: From Stem Cell to Adipocyte , 2005, Critical reviews in biochemistry and molecular biology.

[32]  B. Wolfe,et al.  Treating diabetes with surgery. , 2013, JAMA.

[33]  A. Gefen,et al.  Large, but not Small Sustained Tensile Strains Stimulate Adipogenesis in Culture , 2011, Annals of Biomedical Engineering.

[34]  E. Weiderpass,et al.  Obesity and diabetes epidemics: cancer repercussions. , 2008, Advances in experimental medicine and biology.

[35]  Natan T. Shaked,et al.  Quantitative Analysis of Biological Cells Using Digital Holographic Microscopy , 2011 .

[36]  A. Diaspro,et al.  AFM measurement of the stiffness of layers of agarose gel patterned with polylysine , 2010, Microscopy research and technique.

[37]  Farshid Guilak,et al.  Viscoelastic properties of human mesenchymally-derived stem cells and primary osteoblasts, chondrocytes, and adipocytes. , 2008, Journal of biomechanics.

[38]  R. Burgkart,et al.  Viscoelastic properties of the cell nucleus. , 2000, Biochemical and biophysical research communications.

[39]  Jan Lammerding,et al.  Nuclear mechanics during cell migration. , 2011, Current opinion in cell biology.

[40]  B. Slavin Fine structural studies on white adipocyte differentiation , 1979, The Anatomical record.

[41]  A. Gefen,et al.  Confocal-based cell-specific finite element modeling extended to study variable cell shapes and intracellular structures: the example of the adipocyte. , 2011, Journal of biomechanics.

[42]  H. Green,et al.  An established preadipose cell line and its differentiation in culture II. Factors affecting the adipose conversion , 1975, Cell.

[43]  J. Lammerding,et al.  Biophysical assays to probe the mechanical properties of the interphase cell nucleus: substrate strain application and microneedle manipulation. , 2011, Journal of visualized experiments : JoVE.

[44]  A. Gefen,et al.  Static mechanical stretching accelerates lipid production in 3T3-L1 adipocytes by activating the MEK signaling pathway. , 2012, American journal of physiology. Cell physiology.