Cell contractility arising from topography and shear flow determines human mesenchymal stem cell fate

Extracellular matrix (ECM) of the human Mesenchymal Stem Cells (MSCs) influences intracellular tension and is known to regulate stem cell fate. However, little is known about the physiological conditions in the bone marrow, where external forces such as fluid shear stress, apart from the physical characteristics of the ECM, influence stem cell response. Here, we hypothesize that substrate topography and fluid shear stress alter the cellular contractile forces, influence the genetic expression of the stem cells and hence alter their lineage. When fluid shear stress was applied, human MSCs with higher contractility (seeded on 1 μm wells) underwent osteogenesis, whereas those with lower contractility (seeded on 2 μm gratings) remained multipotent. Compared to human MSCs seeded on gratings, those seeded on wells exhibited altered alignment and an increase in the area and number of focal adhesions. When actomyosin contractility was inhibited, human MSCs did not exhibit differentiation, regardless of the topographical feature they were being cultured on. We conclude that the stresses generated by the applied fluid flow impinge on cell contractility to drive the stem cell differentiation via the contractility of the stem cells.

[1]  J. Emsley,et al.  Integrin connections to the cytoskeleton through talin and vinculin. , 2008, Biochemical Society transactions.

[2]  S. Badylak,et al.  A perivascular origin for mesenchymal stem cells in multiple human organs. , 2008, Cell stem cell.

[3]  S. Cowin,et al.  A model for the excitation of osteocytes by mechanical loading-induced bone fluid shear stresses. , 1994, Journal of biomechanics.

[4]  A. Friedenstein,et al.  STROMAL CELLS RESPONSIBLE FOR TRANSFERRING THE MICROENVIRONMENT OF THE HEMOPOIETIC TISSUES: Cloning In Vitro and Retransplantation In Vivo , 1974, Transplantation.

[5]  Sungho Jin,et al.  Stem cell fate dictated solely by altered nanotube dimension , 2009, Proceedings of the National Academy of Sciences.

[6]  Lin Gao,et al.  Stem Cell Shape Regulates a Chondrogenic Versus Myogenic Fate Through Rac1 and N‐Cadherin , 2010, Stem cells.

[7]  Jiandong Ding,et al.  Effects of aspect ratios of stem cells on lineage commitments with and without induction media. , 2013, Biomaterials.

[8]  Kam W Leong,et al.  Synthetic nanostructures inducing differentiation of human mesenchymal stem cells into neuronal lineage. , 2007, Experimental cell research.

[9]  Benjamin Geiger,et al.  Focal Contacts as Mechanosensors Externally Applied Local Mechanical Force Induces Growth of Focal Contacts by an Mdia1-Dependent and Rock-Independent Mechanism , 2001 .

[10]  Choon Kiat Lim,et al.  Nanotopography modulates mechanotransduction of stem cells and induces differentiation through focal adhesion kinase. , 2013, ACS nano.

[11]  Keng-Hwee Chiam,et al.  Anisotropic rigidity sensing on grating topography directs human mesenchymal stem cell elongation , 2013, Biomechanics and Modeling in Mechanobiology.

[12]  D. Stamenović,et al.  Modulation of Cellular Mechanics during Osteogenic Differentiation of Human , 2004 .

[13]  Rong Fan,et al.  Nanotopography influences adhesion, spreading, and self-renewal of human embryonic stem cells. , 2012, ACS nano.

[14]  S. Sen,et al.  Matrix Elasticity Directs Stem Cell Lineage Specification , 2006, Cell.

[15]  Christoph Ballestrem,et al.  Vinculin controls focal adhesion formation by direct interactions with talin and actin , 2007, The Journal of cell biology.

[16]  Christopher R Jacobs,et al.  Mechanically induced osteogenic differentiation – the role of RhoA, ROCKII and cytoskeletal dynamics , 2009, Journal of Cell Science.

[17]  W. Ostertag,et al.  Proteoglycan synthesis in haematopoietic cells: isolation and characterization of heparan sulphate proteoglycans expressed by the bone-marrow stromal cell line MS-5. , 1997, The Biochemical journal.

[18]  Milan Mrksich,et al.  Geometric cues for directing the differentiation of mesenchymal stem cells , 2010, Proceedings of the National Academy of Sciences.

[19]  C. Lim,et al.  Force-dependent vinculin binding to talin in live cells: a crucial step in anchoring the actin cytoskeleton to focal adhesions. , 2014, American journal of physiology. Cell physiology.

[20]  Michael P. Sheetz,et al.  Stretching Single Talin Rod Molecules Activates Vinculin Binding , 2009, Science.

[21]  N. Gadegaard,et al.  Nanoscale surfaces for the long-term maintenance of mesenchymal stem cell phenotype and multipotency. , 2011, Nature materials.

[22]  L Ploux,et al.  The interaction of cells and bacteria with surfaces structured at the nanometre scale. , 2010, Acta biomaterialia.

[23]  A. Friedenstein.  Stromal mechanisms of bone marrow: cloning in vitro and retransplantation in vivo. , 1980, Haematology and blood transfusion.

[24]  Hanry Yu,et al.  Mechanotransduction In Vivo by Repeated Talin Stretch-Relaxation Events Depends upon Vinculin , 2011, PLoS biology.

[25]  Andrés J. García,et al.  Contractility modulates cell adhesion strengthening through focal adhesion kinase and assembly of vinculin‐containing focal adhesions , 2010, Journal of cellular physiology.

[26]  Lu Wang,et al.  Engineering of a microfluidic cell culture platform embedded with nanoscale features. , 2011, Lab on a chip.

[27]  Christopher S. Chen,et al.  Cell shape, cytoskeletal tension, and RhoA regulate stem cell lineage commitment. , 2004, Developmental cell.

[28]  G. Sukhikh,et al.  Mesenchymal Stem Cells , 2002, Bulletin of Experimental Biology and Medicine.

[29]  M. Stevens,et al.  Changes in embryonic stem cell colony morphology and early differentiation markers driven by colloidal crystal topographical cues. , 2012, European cells & materials.

[30]  M. Riehle,et al.  Interaction of animal cells with ordered nanotopography. , 2002, IEEE transactions on nanobioscience.

[31]  K. Hörmann,et al.  Integrin expression in stem cells from bone marrow and adipose tissue during chondrogenic differentiation. , 2008, International journal of molecular medicine.

[32]  G. Klein.  The extracellular matrix of the hematopoietic microenvironment , 1995, Experientia.

[33]  C. Wilkinson,et al.  The control of human mesenchymal cell differentiation using nanoscale symmetry and disorder. , 2007, Nature materials.

[34]  R. Das,et al.  Influence of nanohelical shape and periodicity on stem cell fate. , 2013, ACS nano.