Revealing elasticity of largely deformed cells flowing along confining microchannels

Deformability is a hallmark of malignant tumor cells. Characterizing cancer cell deformation can reveal how cancer cell metastasizes through tiny gaps in tissues. However, many previous reports only focus on the cancer cell behaviors under small deformation regimes, which may not be representative for the behaviors under large deformations as in the in vivo metastatic processes. Here, we investigate a wide range of cell elasticity using our recently developed confining microchannel arrays. We develop a relation between the elastic modulus and cell shape under different deformation levels based on a modified contact theory and the hyperelastic Tatara theory. We demonstrate good agreements between the model prediction and experimental results. Strikingly, we discover a clear ‘modulus jump’ of largely deformed cells compared to that of small deformed cells, offering further biomechanical properties of the cells. Likely, such a modulus jump can be considered as a label-free marker reflecting the elasticity of intracellular components including the nucleus during cell translocation in capillaries and tissue constrictions. In essence, we perform cell classification based on the distinct micromechanical properties of four cell lines, i.e. one normal cell line (MCF-10A) and three cancer cell lines (MCF-7, MDA-MB-231 and PC3) and achieved reasonable efficiencies (efficiency >65%). Finally, we study the correlation between large-deformational elasticity and translocation rates of the floating cells in the microchannels. Together, our results demonstrate the quantitative analysis of the biomechanical properties of single floating cells, which provide an additional label-free physical biomarker toward more effective cancer diagnosis.

[1]  Byungkyu Kim,et al.  Separation of malignant human breast cancer epithelial cells from healthy epithelial cells using an advanced dielectrophoresis-activated cell sorter (DACS) , 2009, Analytical and bioanalytical chemistry.

[2]  Christopher Beadle,et al.  The role of myosin II in glioma invasion of the brain. , 2008, Molecular biology of the cell.

[3]  Sean X. Sun,et al.  Volume regulation and shape bifurcation in the cell nucleus , 2015, Journal of Cell Science.

[4]  Vera C. Fonseca,et al.  Cellular mechanical properties reflect the differentiation potential of adipose-derived mesenchymal stem cells , 2012, Proceedings of the National Academy of Sciences.

[5]  D. Boal,et al.  Simulations of the erythrocyte cytoskeleton at large deformation. II. Micropipette aspiration. , 1998, Biophysical journal.

[6]  Guangyu Liu,et al.  Multiparametric Biomechanical and Biochemical Phenotypic Profiling of Single Cancer Cells Using an Elasticity Microcytometer. , 2016, Small.

[7]  W. Marsden I and J , 2012 .

[8]  Thomas Lecuit,et al.  Nuclear mechanics in differentiation and development. , 2011, Current opinion in cell biology.

[9]  Millard F. Beatty,et al.  Topics in Finite Elasticity: Hyperelasticity of Rubber, Elastomers, and Biological Tissues—With Examples , 1987 .

[10]  G. Naumov,et al.  Critical steps in hematogenous metastasis: an overview. , 2001, Surgical oncology clinics of North America.

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

[12]  Lap Man Lee,et al.  A microfluidic pipette array for mechanophenotyping of cancer cells and mechanical gating of mechanosensitive channels. , 2015, Lab on a chip.

[13]  G. Piaggio,et al.  Prognostic role of NF-YA splicing isoforms and Lamin A status in low grade endometrial cancer , 2016, Oncotarget.

[14]  Richard S. Chadwick,et al.  Determination of the elastic moduli of thin samples and adherent cells using conical AFM tips , 2012, Nature nanotechnology.

[15]  H. Klocker,et al.  Lamin A/C protein is overexpressed in tissue-invading prostate cancer and promotes prostate cancer cell growth, migration and invasion through the PI3K/AKT/PTEN pathway. , 2012, Carcinogenesis.

[16]  Lidong Qin,et al.  Microfluidics separation reveals the stem-cell–like deformability of tumor-initiating cells , 2012, Proceedings of the National Academy of Sciences.

[17]  Richard Superfine,et al.  Mechanical stiffness grades metastatic potential in patient tumor cells and in cancer cell lines. , 2011, Cancer research.

[18]  Susan C. Roberts,et al.  Pluronic F127 as a cell encapsulation material: utilization of membrane-stabilizing agents. , 2005, Tissue engineering.

[19]  Milica Radisic,et al.  In situ mechanical characterization of the cell nucleus by atomic force microscopy. , 2014, ACS nano.

[20]  Daniele Zink,et al.  Nuclear structure in cancer cells , 2004, Nature Reviews Cancer.

[21]  H. Abdi,et al.  Principal component analysis , 2010 .

[22]  U. Keyser,et al.  Real-time deformability cytometry: on-the-fly cell mechanical phenotyping , 2015, Nature Methods.

[23]  Thibault P. Prevost,et al.  Biomechanics of single cortical neurons. , 2010, Acta biomaterialia.

[24]  Dino Di Carlo,et al.  Hydrodynamic stretching of single cells for large population mechanical phenotyping , 2012, Proceedings of the National Academy of Sciences.

[25]  R. Vink,et al.  Walker 256 tumour cells increase substance P immunoreactivity locally and modify the properties of the blood–brain barrier during extravasation and brain invasion , 2012, Clinical & Experimental Metastasis.

[26]  S. Shapshay,et al.  Detection of preinvasive cancer cells , 2000, Nature.

[27]  E. van den Berg,et al.  Loss of lamin A/C expression in stage II and III colon cancer is associated with disease recurrence. , 2011, European journal of cancer.

[28]  A. Puisieux,et al.  Metastasis: a question of life or death , 2006, Nature Reviews Cancer.

[29]  Ehsan Gazi,et al.  Measurement of elastic properties of prostate cancer cells using AFM. , 2008, The Analyst.

[30]  Joseph J O'Hagan,et al.  Measurement of the hyperelastic properties of 44 pathological ex vivo breast tissue samples , 2009, Physics in medicine and biology.

[31]  H. Shum,et al.  Capillary micromechanics for core-shell particles. , 2014, Soft matter.

[32]  Subra Suresh,et al.  Large deformation of living cells using laser traps , 2004 .

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

[34]  J. Irianto,et al.  Nuclear lamin stiffness is a barrier to 3D migration, but softness can limit survival , 2014, The Journal of cell biology.

[35]  Raymond H. W. Lam,et al.  Microfluidic biosensing of viscoelastic properties of normal and cancerous human breast cells , 2017, 2017 IEEE 12th International Conference on Nano/Micro Engineered and Molecular Systems (NEMS).

[36]  Jeffrey J. Fredberg,et al.  Cytoskeletal stiffness, friction, and fluidity of cancer cell lines with different metastatic potential , 2012, Clinical & Experimental Metastasis.

[37]  R. Jain,et al.  Intratumoral lymphatic vessels: a case of mistaken identity or malfunction? , 2002, Journal of the National Cancer Institute.

[38]  D Needham,et al.  Time-dependent recovery of passive neutrophils after large deformation. , 1991, Biophysical journal.

[39]  Falk Wottawah,et al.  Oral cancer diagnosis by mechanical phenotyping. , 2009, Cancer research.

[40]  Method for measurement of friction forces on single cells in microfluidic devices , 2012 .

[41]  Bjørn Tore Gjertsen,et al.  Axl is an essential epithelial-to-mesenchymal transition-induced regulator of breast cancer metastasis and patient survival , 2009, Proceedings of the National Academy of Sciences.

[42]  E. Nauman,et al.  Mapping nanomechanical properties of live cells using multi-harmonic atomic force microscopy. , 2011, Nature nanotechnology.

[43]  Kyriacos A Athanasiou,et al.  In situ mechanical properties of the chondrocyte cytoplasm and nucleus. , 2009, Journal of biomechanics.

[44]  Jongyoon Han,et al.  Ultra-fast, label-free isolation of circulating tumor cells from blood using spiral microfluidics , 2015, Nature Protocols.

[45]  Y. Tatara On Compression of Rubber Elastic Sphere Over a Large Range of Displacements—Part 1: Theoretical Study , 1991 .

[46]  C. Lim,et al.  Mechanics of the human red blood cell deformed by optical tweezers , 2003 .

[47]  P. Friedl,et al.  Tumour-cell invasion and migration: diversity and escape mechanisms , 2003, Nature Reviews Cancer.

[48]  J. Lammerding,et al.  Nuclear Mechanics and Mechanotransduction in Health and Disease , 2013, Current Biology.

[49]  P. A. van den Brandt,et al.  Lamin A/C Is a Risk Biomarker in Colorectal Cancer , 2008, PloS one.

[50]  Stefan Schinkinger,et al.  Optical deformability as an inherent cell marker for testing malignant transformation and metastatic competence. , 2005, Biophysical journal.

[51]  Masoud Agah,et al.  The effects of cancer progression on the viscoelasticity of ovarian cell cytoskeleton structures. , 2012, Nanomedicine : nanotechnology, biology, and medicine.

[52]  Chwee Teck Lim,et al.  Versatile label free biochip for the detection of circulating tumor cells from peripheral blood in cancer patients. , 2010, Biosensors & bioelectronics.

[53]  Kristian Pietras,et al.  High interstitial fluid pressure — an obstacle in cancer therapy , 2004, Nature Reviews Cancer.

[54]  J. Rao,et al.  Nanomechanical analysis of cells from cancer patients. , 2007, Nature nanotechnology.